Research Paper Volume 11, Issue 16 pp 6490—6502

Figure 3. Hair cells were vulnerable to ROS injury after Cav1.3 was knocked out. (A) the effect of CaV1.3 knock out in HEI-OC1 was analyzed by flow cytometry. (B) membrane potential (top) and non-linear capacitance (NLC) (bottom) studies in WT HEI-OC1 and CaV1.3 KO HEI-OC1 cells (n=5). (C) western-blotting analysis of CaV1.3 and p53 expression in control and senescence HEI-OC1 cells induced by D-galactose (D-Gal) or hydrogen peroxide (H₂O₂), the bottom panel is the quantitative analysis. (D) β-Galactosidase staining (top) and C12FDG staining (bottom) of control and senescent HEI-OC1 cells induced by H₂O₂. (E) flow cytometry analysis of CaV1.3 in control and H₂O₂ induced HEI-OC1 cells. (F) C12FDG staining (top) and β-Galactosidase staining (bottom) of NC (negative control) and KO (CaV1.3 knock out) HEI-OC1 cells after H₂O₂ induction. (G, H) CFSE staining and red dot staining of NC and KO HEI-OC1 cells with or without H₂O₂ induction. (I) LDH assay of NC and KO HEI-OC1 cells after H₂O₂ induction (n=3). (J) caspase-3/7-AAD staining of NC and KO HEI-OC1 cells after H₂O₂ induction (n=3). Error bars represent mean ± s.d.; *P<0.05; **P < 0.01; ***P < 0.001; n.s. not significant; by one-way analysis of variance (ANOVA) (I, J).