Figure S4. SV morphology and EP are... | Rockefeller University Press

Figure S4.

$SV morphology and EP are normal in Tprn−/−mouse. (A) EP of P60 WT (Tprn+/+, two males, one female), heterozygous (Tprn+/−, two males, three females), and homozygous (Tprn−/−, three males, three females) mutant mice was within the WT range and indistinguishable between all three genotypes. Only left ears were tested in all three genotypes. (B and C) The inner ears of EP tested mice were immunostained, and the percentage of OHC and IHC loss was quantified and calculated as a fraction of number of missing hair cells out of the total number of this cell type in all images of middle and apical turn of the same ear. Error bars represent SD. One ear from each mouse was imaged and analyzed. Total of three animals were used for each genotype group. (B)Tprn−/− mice had higher percentage of OHC loss compared with the Tprn+/− and Tprn+/+. (C) No statically significant difference was observed between the percentage of IHC loss of the Tprn−/− mice with respect to the Tprn+/− and Tprn+/+. P values <0.001 were denoted with ***; unpaired two-sided t test. (D) Representative images of OC of EP tested mice of each genotype showing three rows of OHCs and one row of IHCs stained with antibody to MYO7A as a hair cell marker. Phalloidin is used to visualize the F-actin. Locations of missing OHCs are indicated by white arrows. Scale bars, 10 µm. (E) Cross-sections of the SV from the contralateral ear of EP tested animals. Cryosections stained for KCNJ10 (magenta, an intermediate cell marker), DAPI (nuclear marker), and phalloidin (green, F-actin) did not show any differences in SV staining and morphology of the Tprn−/− mice compared with the Tprn+/− and Tprn+/+ littermates. Scale bars 20 µm.$

SV morphology and EP are normal in Tprn ^−/− mouse. (A) EP of P60 WT (Tprn^+/+, two males, one female), heterozygous (Tprn^+/−, two males, three females), and homozygous (Tprn^−/−, three males, three females) mutant mice was within the WT range and indistinguishable between all three genotypes. Only left ears were tested in all three genotypes. (B and C) The inner ears of EP tested mice were immunostained, and the percentage of OHC and IHC loss was quantified and calculated as a fraction of number of missing hair cells out of the total number of this cell type in all images of middle and apical turn of the same ear. Error bars represent SD. One ear from each mouse was imaged and analyzed. Total of three animals were used for each genotype group. (B)Tprn^−/− mice had higher percentage of OHC loss compared with the Tprn^+/− and Tprn^+/+. (C) No statically significant difference was observed between the percentage of IHC loss of the Tprn^−/− mice with respect to the Tprn^+/− and Tprn^+/+. P values <0.001 were denoted with ***; unpaired two-sided t test. (D) Representative images of OC of EP tested mice of each genotype showing three rows of OHCs and one row of IHCs stained with antibody to MYO7A as a hair cell marker. Phalloidin is used to visualize the F-actin. Locations of missing OHCs are indicated by white arrows. Scale bars, 10 µm. (E) Cross-sections of the SV from the contralateral ear of EP tested animals. Cryosections stained for KCNJ10 (magenta, an intermediate cell marker), DAPI (nuclear marker), and phalloidin (green, F-actin) did not show any differences in SV staining and morphology of the Tprn^−/− mice compared with the Tprn^+/− and Tprn^+/+ littermates. Scale bars 20 µm.