To screen for the best-performing siRNA of each group, we employed a dual-luciferase assay-based system. The respective target sequences were individually inserted into the 3′ UTR of a plasmid-located Renilla luciferase

gene. The DNA polymerase, pTP, IVa2, hexon, and protease siRNAs, together with http://www.selleckchem.com/products/gsk1120212-jtp-74057.html the respective reporter vectors, were used to co-transfect HEK293 cells. Knockdown of Renilla luciferase expression in relation to the expression of a firefly luciferase gene located on the same plasmid was determined in dual-luciferase assays. The silencing capacity of the E1A siRNAs was assessed in A549 cells because promoter activities of the reporter vectors turned out to be altered upon silencing of the endogenous E1A gene present in HEK293 cells ( Graham et al., 1977) (data not shown). For all target mRNAs,

we identified siRNAs enabling a knockdown of ⩾78% at a concentration of 30 nM ( Fig. 1). The best-performing siRNAs of each group, i.e., pTP-si8, Pol-si2, Hex-si2, E1A-si3, Iva2-si2, and Prot-si1, were selected for further characterization. The dual-luciferase assay-based screening system was employed to select the best-performing siRNAs of each group. Next, we investigated whether the selected siRNAs were able to knockdown gene expression during an adenovirus infection of A549 cells. Cells were transfected with the siRNAs at a concentration of 10 nM, and then infected with Ad5 at an MOI of 0.01 TCID50/cell. Target mRNA levels were determined Everolimus price by RT-qPCR, using primers specific for the individual mRNAs (Fig. 2A).

The highest silencing rates (93–97%) were observed for the E1A-, DNA polymerase-, pTP-, and IVa2-targeting Montelukast Sodium siRNAs. Silencing of the hexon and protease genes was less pronounced (79–87%). Except for the difference in residual hexon and pTP mRNA levels, the differences between hexon or protease mRNA levels and those of all other early genes were statistically significant. As the pTP, DNA polymerase, and IVa2 mRNAs share a common 3′ part (Supplementary Fig. 1), and the DNA polymerase target site is also part of the pTP mRNA, IVa2- and DNA polymerase-directed siRNAs were therefore expected to concomitantly silence pTP/DNA polymerase/IVa2 and pTP/DNA polymerase, respectively. Furthermore, siRNA-mediated silencing of early genes was expected indirectly to affect the expression of those middle or late genes for which expression is known to depend on early viral gene products. Thus, we also determined the effect of the E1A-, pTP-, DNA polymerase-, and IVa2-targeting siRNAs on the expression of the other genes. Silencing of E1A resulted in a marked reduction in the expression of all other genes (Fig. 2B). This can be attributed to the central role of E1A in activating the expression of downstream genes. Silencing of the E1A gene actually resulted in a greater reduction in the expression of hexon and protease than did direct silencing of these genes by the hexon and protease siRNAs (compare Fig. 2A and B).

, 2008 and Timms and Moss, 1984).

Another indication of an upcoming shift in this region can be found in the increasing dominance of floating macrophytes at the expense of the submerged Ulixertinib mouse macrophytes (Scheffer et al., 2003 and Zhao et al., 2012b). Floating macrophytes are able to better cope with lower light conditions than submerged macrophytes because they grow at the water surface. When light conditions deteriorate close to the shifting point, floating macrophytes will therefore predominate submerged macrophytes (Scheffer et al., 2003). While macrophytes disappeared, the total primary production of Taihu increased more than twofold from 1960 (5.46 t · km− 2 yr− 1) to 1990 (11.66 t · km− 2 yr− 1) owing to the increasing phytoplankton biomass that bloomed due to the excessive nutrient input (Li et al., 2010). The first algal blooms occurred in 1987 in Meiliang

Selleck GSK2118436 Bay (Fig. 5, 1980s). Subsequently, algal blooms dominated by non-N2 fixing cyanobacteria (Microcystis) increased in coverage and frequency, and appeared earlier in the season ( Chen et al., 2003b, Duan et al., 2009 and Paerl et al., 2011b). The presence of mainly non-N2 fixing cyanobacteria indicates that external and internally-supplied nitrogen are sufficient to maintain proliferation over N2-fixers ( Paerl et al., 2011b). The early blooms in the northern bays and western shores occurred right where enrichment was

most severe and easterly winds drove algae to form thick scums ( Chen et al., 2003b and Li Rucaparib chemical structure et al., 2011a). At that time, high concentrations of suspended solids in the lake centre due to wind action ( Fig. 8) might have prevented algal growth by light limitation ( Li et al., 2011a and Sun et al., 2010). Despite this mechanism, blooms also emerged in the lake centre from 2002 onwards ( Duan et al., 2009). Finally, in 2007 the problems with drinking water became so severe that it was not possible to ignore the blooms anymore ( Qin et al., 2010). The effects of excessive nutrient loads go beyond the shift in primary producers alone and appear also higher in the food web. As the biomass of primary producers and zooplankton grew over time, the biomass of higher trophic levels shrank and several species disappeared (Guan et al., 2011 and Li et al., 2010). There are indications that in the presence of Microcystis, the zooplankton shifted their diet to the detritus-bacteria pathway rather than grazing on living phytoplankton ( de Kluijver et al., 2012). A macroinvertebrate survey in 2007 by Cai et al. (2012) showed that small individuals (e.g. Tubificidae) appear in large numbers in the algal blooming zone ( Fig. 5, 2007). The appearance of mainly small macroinvertebrate species might be related to the absence of refuges to prevent predation (e.g. macrophytes) ( Cai et al.

g. pointbar deposits, deserted channels, and abandoned oxbow lakes), (2) and floodplain cover deposits, formed by vertical accretion of fine sediments in slow-moving floodwaters of the

basins. Cover deposits are widespread along the flanking zone from Jacobabad to Manchar Lake, in the southeast around Mirpur Khas and Umarkot, and in the delta (Holmes, GW3965 in vitro 1968). The historical Indus River sent off distributaries and small seasonal spillway channels toward its flanks and across the delta. Such smaller-scale channels are characterized by levees rather than by river bars and meander scrolls. Levees of the Ghar and Western Nara (Fig. 1) are ∼3 m high due to periodic overspill of their selleck compound banks and define these 3 km-wide paleochannels. Narrower channels and shorter wavelength meanders define former courses of the

Indus: the Khairpur at between 4 km and 8 km; Shahdapur at 5 km; and the Warah at 6 km (Fig. 1). The modern Indus is wider with larger but fewer meanders (∼14 km wavelength). Sinuosity of the paleo-Indus channels (Fig. 1 and Fig. 2) had a range from: (1) Badahri: 1.51, (2) Warah: 1.55, (3) Kandhkot: 1.65; (4) Puran: 1.81, (5) Shahdadkot: 1.99, (6) Eastern Nara: 2.05, (7) Khairpur: 2.33, and (8) Shahdadpur: 2.51. The modern Indus has sinuosity values ranging from 1.1 to 2.0 with a mean value of 1.8 (see discussion below). Paleochannels therefore had similar or sometimes greater sinuosity. The visible record of paleochannels represents only the last ∼1000 years. The remotely sensed topography of Fig. 2 perhaps captures some of the longer record of river avulsion and floodplain development and demonstrates how the floodplain aggrades through major avulsions of the trunk Indus. The large channel belt switches leaving behind 1–3 m of super-elevated channel belt deposits that shed crevasse-splay fingers

and fans interweaving with cover deposits to their sides (Fig. 2, Fig. 3, Fig. 4 and Fig. 5). An interesting feature of the imaged floodplain topography is its fan-like appearance (Fig. 2 and Fig. 5). When viewed along valley profiles (Fig. 3), these fan-like waves have a first order wavelength of 29 km, upon which is superimposed a second Arachidonate 15-lipoxygenase order set of waveforms with wavelength of ∼3.6 km. We suggest that the first order waveform reflect the avulsion frequency of the main Indus River (on the order of several centuries). Major avulsions shift the loci of floodplain deposition suddenly, leaving behind these first-order super-elevated fan lobes (see Fig. 2B). Whereas the second-order scale features perhaps relate to decadal occurrence of floods that build up intermingled crevasse deposits around the larger paleochannel features (Fig. 5). The width and depth of the modern Indus and other paleochannels are well demonstrated in both strike sections (Fig. 4) and plan view (Fig. 5).

Prehistoric animals likely did not attain significantly greater depths; dinosaur burrows, for example, were long unrecorded, and the single example known ( Varricchio et al., 2007) is not much more than 20 cm across and

lies less than a metre below the palaeo-land surface. Plant roots can penetrate depths an order of magnitude greater, especially in arid regions: up to 68 m for Boscia truncata in the Kalahari desert ( Jennings, 1974). They can be preserved as rootlet traces, generally through diagenetic mineral precipitation or remnant carbon traces. Roots, though, typically infiltrate between sediment grains, limiting the amount of sediment displacement and hence disruption to the rock fabric. selleck kinase inhibitor At a microscopic level, too, there is a ‘deep biosphere’ composed of sparse, very slowly metabolizing microbial communities that can exist in pore spaces and rock fractures to depths of 1–2 km (e.g. Parkes et al., 1994). These may mediate diagenetic reactions where concentrations

of nutrients allow larger populations (such as the ‘souring’ of oil reservoirs) but otherwise leave little trace in the rock fabric. Very rarely, these communities have been found to be accompanied by very deep-living nematode worms (Borgonie GDC-0973 in vitro et al., 2011), but these seem not to affect the rock fabric, and we know of no reports of their fossil remains or any traces made by them. The extensive, large-scale disruption of underground rock fabrics, to depths of >5 km, by a single biological species, thus represents a major geological innovation (cf. Williams et al., 2014). It has no analogue in the Earth’s 4.6 billion year history, and possesses some sharply distinctive features: for instance, the structures produced reflect a wide variety of human behaviour effected through tools or more typically mechanized excavation, rather than through bodily activity. Hence, the term ‘anthroturbation’ (Price et al., 2011; see also Schaetzl

and Anderson, 2005 for use in soil terminology) is fully justified, and we use this in subsequent description below. This is extensive, HSP90 and distantly analogous to surface traces left by non-human organisms. It includes surface excavations (including quarries) and constructions, and alterations to surface sedimentation and erosion patterns, in both urban and agricultural settings. Its nature and scale on land has been documented (e.g. Hooke, 2000, Hooke et al., 2012, Wilkinson, 2005, Price et al., 2011 and Ford et al., 2014) and it extends into the marine realm via deep-sea trawling (e.g. Puig et al., 2012) and other submarine constructions. Here we simply note its common presence (Hooke et al.

On the other hand, new civil protection challenges arise in localized areas and periods

of the year, from an increasing pressure brought by mountain tourism. Preparedness is becoming Crenolanib chemical structure a core issue where the wildland–urban interface is being expanded, and new strategies have to be considered, along with actual impacts of fires on the ecosystem services, especially within the perspective of integrating fire and erosion risk management. We gratefully acknowledge the Joint Research Centre, European Commission, for providing forest fires data (yearly burnt area) accessible from the European Forest Fire Information System (EFFIS). They have been used for calculating statistics about the incidence of forest fires in the Alpine SCH772984 solubility dmso region during last decades. “
“In 2003, an editorial in the journal Nature ( Nature editorial, 2003) proclaimed that human activity has created an Anthropogenic Earth, and that we now lived in the Anthropocene, an epoch where human–landscape interactions alter the Earth morphology, ecosystems and processes ( Ellis, 2011, Zalasiewicz et al., 2008, Zalasiewicz et al., 2011, Tarolli et al., 2013, Tarolli, 2014, Tarolli et al., 2014a and Tarolli et al., 2014b). One of the most important human domination of land systems is the creation of the reclamation and drainage networks that have a key role in agricultural and environmental sustainability, and can transform

landscapes and shape history ( Earle and Doyle, 2008). Following the land-use changes, drainage networks faced deep alterations due to urbanization and soil consumption ( Cazorzi et al., 2013), but also due to demographic pressure ( Fumagalli, 1976, Hallam, 1961 and Millar and Hatcher, 1978),

and changes in technological innovation ( Magnusson, 2001 and van Dam, 2001), and agricultural techniques. At the same time drainage networks faced an under-investment in their provision and maintenance ( Scheumann and Freisem, 2001) with insufficient evacuation of water runoff in large parts of the reclaimed areas ( Curtis and Campopiano, 2012), and they became crucial in the control of flood generations ( Gallart et al., 1994, Voltz et al., 1998, Marofi, 1999, Moussa et al., 2002, Evrard et al., 2007, Pinter et al., 2006, Bronstert et al., 2001, Pfister et al., 2004, Savenije, Uroporphyrinogen III synthase 1995, Wheater, 2006 and Palmer and Smith, 2013). In earlier times and with less available technology, land drainage and land use was largely determined by the function that could be performed by the natural soil. However, in the course of the last century this relation between soil draining functions and land use has been lost to a certain extent ( Scalenghe and Ajmone-Marsan, 2009), and numerous researches underlined how land use changes altered the local hydrological characteristics ( Bronstert et al., 2001, Brath et al., 2006, Camorani et al., 2005, Heathwaite et al., 1989, Heathwaite et al.

7% per cm; and for fish with 4% learn more lipid, the rate was 2.1% per cm. Coho with high filet % lipid exhibited higher PCB concentrations even at small lengths, but PCB concentrations appeared to increase at a slower rate in these fish as length increased. While these interactions improved the fit of the model, they represent only minor changes in the primary relationships among PCB concentrations and time, body length, % lipid, and season that were suggested by the original main effects model

described previously. Exploratory plots and GAM models suggested patterns for chinook similar to coho with a rapid decline in filet PCB concentrations until the mid to late 1980s, then a slower decline to the 2010; increases in PCB concentrations as both body length and % lipid in filets increased; and higher PCB concentrations Obeticholic Acid molecular weight in filets from fish collected in the fall than in the summer. We fit the same set of models

that we fit for coho, and estimated the point of intersection of piecewise linear trends to be 1985, one year later than for coho. The two models for chinook with lowest AIC included the same predictors as the two best-fitting models for coho: predictors for the model with minimum AIC were piecewise linear time trends, fish body length, % filet lipid, and season collected (Table 4). The model including the additional predictor of location fit slightly worse. The estimated rate of decrease in PCB concentration was − 16.7% per year for 1976–1985 (95% CI: − 18.2% to − 15.2%) and − 4.0% per year for 1986–2010 (95% CI: − 4.4% to − 3.6%; Table 5 and Fig. 3). PCB concentration increased by 2.3% per cm of length (95% CI: 2.1% to 2.5%) and by 10.2% for each 1% increase in % lipid (95% CI: 8.9% to 11.6%). For chinook at a given length and % lipid content, PCB concentrations were 80.6% larger for fish caught in the fall than the summer (95% CI: 67.7% to 94.5%). As with coho, we also examined models that included condition as a predictor using a smaller dataset containing only records with condition. Similar to our findings

with coho, models with minimum AIC were the same as those for the larger dataset; models including condition fit substantially worse. We examined models with all combinations of 2-way interactions among the predictor variables in the model just described; among those models, the one with minimum AIC included 2-way interactions between chinook body Clostridium perfringens alpha toxin length and the two time trends, between length and season, and between length and % lipid. The interactions between body length and the time trends suggested that larger chinook exhibited slower declines than smaller fish in the early time period (− 17.7% for a 60 cm fish vs − 13.3% for a 100 cm fish), but more rapid declines in the later time period (− 3.5% for a 60 cm fish vs − 5.3% for a 100 cm fish). The interaction between chinook body length and season caught was due primarily to differences in filet PCB concentrations for smaller fish between the two seasons.

Many bioactive constituents are present in ginseng extracts, and ginsenosides, the main constituents of ginseng, are believed to have antiallergic, antioxidant, and immune-stimulatory activities [3]. The two traditional preparations of Korean ginseng, white ginseng (WG) and red ginseng (RG), are presumed to have different bioactivities in traditional medicine. WG is produced by the sun drying of fresh ginseng, whereas RG is manufactured by steaming fresh

ginseng and then drying it to a moisture content of < 15% [4]. Many researchers have reported that Depsipeptide price the steaming process increases the bioactivity of ginseng [4], [5] and [6]. Few comparative studies have been conducted on the effects of WG Erastin cost and RG on various diseases. Asthma is a serious health problem and affects people of all ages, and its most common trigger is continuous exposure to allergens [7]. Allergic asthma is characterized by increased mucus production, reversible airway obstruction, eosinophil infiltration, and nonspecific airway hyperresponsiveness (AHR) [8]. The development of asthma is mediated by the overexpression of T helper type 2 (Th2)-mediated or Th1-mediated cytokines, such as interleukin (IL)-4, IL-5, etc. [8] and [9]. However, currently available therapies cannot completely

control the symptoms of asthma, and even intensive treatment shows little effect on healthcare utilization [10]. Consequently, efforts are

required to identify new remedies, preferably of natural origin, for mitigating the effects of these immune-related disorders. P. ginseng is one of most commonly used medicinal herbs to complement the treatment of asthma, allergies, and immunologic conditions [11]. Several researchers have reported that P. ginseng ameliorates asthma in animal models [12] and [13], but to date, the effects of processing on its medicinal effects have not been studied. Therefore, in the present study, we compared the effects of Casein kinase 1 WG and RG in a mouse model of acute asthma. In previous studies we reported that herbal remedies offer potential complementary or alternative treatments and showed that the regulation of Th1/Th2 balance could provide a strategy for the treatment of respiratory diseases [14] and [15]. In this study, we investigated the effects of WG and RG on the infiltration of inflammatory cells, on airway remodeling, and on expressions of inflammation-related cytokines in an ovalbumin (OVA)-sensitized mouse model of acute asthma. Seven-week-old female BALB/c mice (Daehan Biolink, Chungbuk, Korea) were housed in polypropylene cages at 24 ± 4°C under a 12 h light and dark cycle for at least 1 week prior to experiments. Animals were fed with a standard pellet diet and supplied water ad libitum.

Once seen as the margins of our

planet (see Kirch, 1997), islands have emerged as centers of early human interaction, demographic expansion, and exploration (Erlandson and Fitzpatrick, 2006, Rainbird, 2007 and Fitzpatrick and Anderson, 2008). Islands are important both as microcosms of the patterns and processes operating on continents and as distinct locations with often greater isolation and unique biodiversity. Data from the Americas, Australia, Southeast Asia, the Pacific, North Atlantic, Mediterranean, and Caribbean demonstrate a deep history of maritime voyaging that suggests that for anatomically modern humans (Homo sapiens), the ocean was often a pathway of human interaction and discovery rather than a major obstacle or barrier

selleck inhibitor ( Anderson et al., 2010a, Erlandson, 2001, Erlandson, 2010a and Erlandson, 2010b). In other cases, ocean currents, winds, and other processes can influence travel across the waters surrounding islands ( Fitzpatrick and Anderson, 2008 and Fitzpatrick, 2013). Understanding when humans first occupied islands is important for understanding the geography and ramifications of ancient human environmental interactions. Here we outline

the antiquity of island colonization in major island groups around the world to contextualize our Metformin molecular weight discussion of Polynesia, the Caribbean, and California. The earliest evidence for island colonization by hominins may be from Flores in Southeast Asia, which appears to have been colonized by Homo erectus 800,000 or more years ago ( Morwood et al., 1998 and Morwood find more et al., 2004). Evidence for maritime voyaging and island colonization is very limited, however, until after anatomically modern humans spread out of Africa about 70,000–60,000 years ago ( Erlandson, 2010a and Erlandson, 2010b). Australia and New Guinea were colonized roughly 45,000–50,000 years ago ( O’Connell et al., 2010 and O’Connor, 2010) in migrations requiring multiple sea voyages up to 80–90 km long. Several island groups in Southeast Asia were also settled between about 45,000 and 30,000 years ago, and some of these early maritime peoples appear to have had significant marine fishing capabilities ( O’Connor, 2010 and O’Connor et al., 2011). Additional long sea voyages were required for humans to colonize the Bismarck Archipelago in western Melanesia between 40,000 and 35,000 years ago ( Erlandson, 2010a).

Histological examination of thoracic flight muscle in these flies revealed evidence of pronounced myopathy in flies expressing mutant dVCP, including CDK inhibitor atrophy of individual muscles and loss of normal sarcomere architecture ( Figure 1F). Ultrastructural examination of muscle tissue by transmission electron microscopy (TEM) revealed

marked morphological abnormalities in mitochondria with extensive megaconia and pleioconia ( Figure 1F). Interestingly, prior phenotypic analysis of flies expressing mutant VCP reported that degeneration was accompanied by reduced cellular ATP levels ( Chang et al., 2011). The mechanism of altered ATP levels was not explored in Chang et al. Nevertheless, the relevance of the altered ATP levels was

nicely demonstrated Bcl 2 inhibitor since artificial manipulation of ATP levels modified the degenerative phenotype ( Chang et al., 2011). The myopathy and specific mitochondrial abnormalities observed in dVCP mutant flies are reminiscent of the phenotypes reported in flies null for PINK1 and Parkin (Greene et al., 2003; Poole et al., 2008). Our interest in a possible connection to these genes was heightened by the fact that a subset of patients with VCP mutations present with parkinsonism or Parkinson’s disease (Kimonis et al., 2008; Spina et al., 2013), a clinical phenotype also associated with mutations in PINK1 and Parkin. PINK1 and Parkin participate in a common pathway that regulates mitochondrial dynamics and serve to maintain mitochondrial quality control (Clark et al., 2006; Narendra et al., 2008, 2010; Park et al., 2006). These observations led us to hypothesize that VCP might be a component of the PINK1/Parkin pathway and contribute to mitochondrial quality control. To test this hypothesis, we performed epistasis studies Niclosamide between VCP, PINK1, and Parkin. We determined that overexpression of VCP rescued the degenerative phenotype associated with PINK1 deficiency, as evidenced by suppression of thoracic indentations ( Figures 2A and 2B) and restoration

of normal locomotor function ( Figure 2C) in PINK1 null (PINK1B9). Furthermore, histological analysis demonstrated that VCP overexpression rescued the mitochondrial phenotype in PINK1 null flies ( Figure 2D). This rescue by VCP is similar to that observed by overexpressing Parkin in PINK1 null flies ( Clark et al., 2006; Park et al., 2006). These results indicate that, like Parkin, VCP functions downstream of PINK1 in the mitochondrial quality-control pathway. In contrast, VCP did not suppress the degenerative phenotype associated with Parkin deficiency ( Figures 2A–2C). These data indicate that VCP functions upstream or in concert with Parkin or, alternatively, independently of Parkin in supporting mitochondrial quality control by PINK1.

In order for the RNA variant to be detected, each 50-mer probe must bind in tandem (schematic shown in Figure 1C). Similar to previous studies, C9ORF72 V1 and V2 RNA levels were attenuated by ∼50% in C9ORF72 ALS CNS tissue (n = 5–10) compared to healthy controls (n = 4–10) (Figure 1D). Interestingly, iPSNs (n = 3) exhibited

a similar expression pattern with an approximate 50% reduction in C9ORF72 V1 and V2 RNA levels, while patient fibroblasts showed low C9ORF72 expression with no obvious differences between the control and C9ORF72 ALS lines (although slight differences may be confounded by the lower absolute RNA counts) (Figure 1D). In human CNS tissue, the highest quantity of C9ORF72 RNA is found in the cerebellum and cervical spinal cord. Importantly, in non-C9ORF72 ALS tissue, C9ORF72 RNA levels were comparable to healthy control tissue check details Saracatinib in vitro (Figure 1E). While we tested for the presence of C9ORF72 V3 RNA using probes targeting the spliced mRNA sequence (Figure 1C), it could only be detected slightly above background in iPSNs and cervical spinal cord tissue with no obvious expression differences between control and

C9ORF72 ALS cells/tissue (Figure S2A). However, probes targeting C9ORF72 intron 7, which detect V2 and V3 pre-mRNA, exhibited a statistically significant reduction of this RNA sequence in C9ORF72 ALS compared to control and non-C9ORF72 ALS tissue, suggesting that the prespliced levels of C9ORF72 V2 and V3 RNA are also reduced in endogenous tissue (Figure S2B). Whether these changes in RNA correlate with endogenous protein are uncertain, as current

antibodies to C9ORF72 protein are not suitable for specific quantification (data not shown). While the function of the C9ORF72 protein is unknown, a recent study in zebrafish has shown that C9ORF72 Flavopiridol (Alvocidib) deficiency via ASO knockdown results in locomotor deficits and abnormal growth and morphology of motor axons (Ciura et al., 2013). Further studies are required to determine whether a C9ORF72 deficit in human ALS/FTD contributes to the disease phenotype. We detected intranuclear GGGGCC RNA foci in C9ORF72 ALS patient tissue, iPSNs, and fibroblasts by RNA fluorescent in situ hybridization (RNA FISH), similar to previous reports of postmortem CNS tissue (DeJesus-Hernandez et al., 2011), employing fluorescently labeled locked nucleic acid (LNA) oligonucleotide probes targeting the expanded repeat (Figures 2A and S3A). This pathology was not present in non-C9ORF72 control cells or C9ORF72 cells labeled with scrambled FISH probes (Figures 2B and S3A). Specifically, we observed that, across five C9ORF72 fibroblast and four C9ORF72 iPSN lines, approximately 25% and 35% of cells contained intranuclear GGGGCC RNA foci, respectively (Figures 2B, 2C, and S3A).