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. 2025 Feb 26;21(5):2313–2329. doi: 10.7150/ijbs.105667

Figure 6.

Figure 6

The protective effects of rIPC on TCMK-1 cells are enhanced by NOX4 silencing. (A) TCMK-1 cells were transfected with negative control (NC) siRNA or NOX4 siRNA for 6 h and then treated with 2 μg/ml cisplatin for 24 h. The knockdown efficiency of NOX4 siRNA in TCMK-1 cells was evaluated by RT-qPCR analysis and western blot analysis. (B) NOX4 expression evaluated by RT-qPCR analysis, western blot analysis and quantified by densitometry. (C) The ROS production in TCMK-1 cells was assessed by DCFH-DA staining (100x, scale bar = 100 μm). (D) Mitochondrial dynamic regulatory molecules (DRP-1, OPA-1 and MFN-2) evaluated by RT-qPCR analysis, western blot analysis and quantified by densitometry. (E) Mitophagy level (PINK1, p62/SQSTM1 and LC3B) measured by RT-qPCR analysis, western blot analysis and quantified by densitometry. (F) Ferroptosis-regulatory molecules (ACSL4 and GPX4) assessed by RT-qPCR analysis, western blot analysis and quantified by densitometry. Data are presented as mean ± SD, n = 3. CCCP: carbonyl cyanide 3-chlorophenylhydrazone, CP: cisplatin, ROS: reactive oxygen species. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.