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. 2021 Jul 10;12(1):167–181. doi: 10.1016/j.apsb.2021.07.002

Figure 6.

Figure 6

Effects of astaxanthine on cisplatin-induced apoptosis in HEI-OC1 cell line and cochlear hair cells. (A) Representative Myosin VIIA (green) and cleaved caspase-3 (red) stained confocal images of postnatal 3-day C57BL/6 mouse cochlear explants from the control, CDDP, AST, and AST plus cisplatin (AST/CDDP) groups. The co-staining of Myosin VIIA and cleaved caspase-3 are identified with the white labels. Scale bars = 20 μm. (B) Quantification of the numbers of cleaved caspase-3 and Myosin VIIA co-staining cells per 200 μm in the cochlea after the different treatments. The data is presented as means ± SD, ∗∗∗∗P < 0.0001 compared with control (blue) and AST/CDDP (black) by two-way analysis of variance (ANOVA) with Bonferroni correction. (C) Representative Myosin VIIA (green) and TUNEL (magenta) stained confocal images of postnatal 3-day C57BL/6 mouse cochlear explants from the control, CDDP, AST, and AST/CDDP groups. The co-staining of Myosin VIIA and TUNEL are identified with white labels. Scale bars = 20 μm. (D) Quantitative analysis of the numbers of Myosin VIIA and TUNEL co-staining cells per 200 μm in the cochleae after the different treatments. The data is presented as means ± SD, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 compared with control (blue) and AST/CDDP (black) by two-way ANOVA with Bonferroni correction. (E) The mRNA expression profiles of apoptotic markers including Bax, caspase-3, caspase-8, caspase-9, and Fadd in HEI-OC1 cells following different treatments. The values of qPCR were normalized to β-actin and averaged, mean ± SD, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 compared with control (blue) and CDDP (black) by the one-way ANOVA with Bonferroni correction, n = 3 for each group. (F) Representative Western blot images of BAX, BCL-2 and cleaved caspase-3 in HEI-OC1 cells from different treatments. All bands of protein were normalized to β-actin and averaged, mean ± SD, ∗∗P < 0.01, ∗∗∗P < 0.001 compared with control (blue) and CDDP (black) by one-way ANOVA with Bonferroni correction, n = 3 for each group. AST, astaxanthine; CDDP, cisplatin; Base, basal turn; Middle, middle turn; Apex, apical turn.