The media was replaced with virus-free MEM supplemented with 2% BSA. Cultures were maintained for an additional 2 to 4 days before recording. Student’s (two-tailed) t test and one-way ANOVA was used to compare ABR Sunitinib price thresholds and biophysical properties of mechanotransduction currents using the statistical function in Origin7.5 (OriginLab). Statistical significance is indicated

as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Data are presented as mean ± 1 standard deviation unless otherwise noted. To analyze single-channel conductances in wild-type cells (Figure 5D) we used three statistical tests, cubic clustering criterion, pseudo F statistics, and pseudo T-squares statistics, which are components of SAS Selleckchem Selumetinib 9.2 software (SAS Institute Inc). All animal

experiments and procedures were performed according to protocols approved by the Animal Care and Use Committee of the National Institute on Deafness and Other Communication Disorders and the Animal Care and Use Committee of Boston Children’s Hospital (Protocol #1959 and #2146). We thank Erin Child for technical assistance, Lin Huang for assistance with statistical analysis, and Martin Hrabé de Angelis and Helmut Fuchs for providing Beethoven mice. We thank John Assad, David Clapham, David Corey, Tom Friedman, Matt Kelley, Tom Schwarz, and Clifford Woolf for helpful discussions and critical review of an earlier version of the manuscript. This work was supported by NIDCD intramural research funds Z01-DC000060-10 (A.J.G.);

NIH grants R01-DC05439 (J.R.H.) and R01-DC008853 (G.S.G.). K.K. and A.J.G. hold US Patents: 7,166,433 (Transductin-2 and Applications to Hereditary Deafness; abandoned in 2009), 7,192,705 (Transductin-1 and Applications to Hereditary Deafness), and 7,659,115 (Nucleic Acid Encoding Human Transductin-1 Polypeptide). These patents have never been licensed and generate no income or royalties. B.P. collected all electrophysiology data from inner hair cells and helped analyze data. G.S.G. collected electrophysiology data from vestibular hair cells and helped Levetiracetam analyze the data. G.C.H. collected electrophysiology data from vestibular hair cells, performed ABRs, and helped analyze data. Y.A. generated adenoviral vectors and performed qRT-PCR experiments. K.K. generated Tmc constructs. K.I. performed cell count analysis of Tmc1 mutant mice. Y.K. acquired SEM images of hair cells. A.J.G. contributed to conception of the study and provided critical comments on the data and manuscript. J.R.H. conceived and designed the experiments, analyzed the data, generated the figures, and wrote the paper. All authors provided comments and approved the final submission of the manuscript. “
“Layer 5B (L5B) pyramidal neurons are the major output neurons of the neocortex, and so represent the final site of neocortical integration.