FIGURE 5.

H₂S protected against RSL3-induced ferroptosis in Sertoli cells. (A–D) Effect of H₂S on RSL3-induced Sertoli cell death. TM4 cells were incubated with 5 µM RSL3 with or without 1 mM NaHS for 24 h (A) or in the presence or absence of 2 mM BCA for 16 h (B). The cell viability was determined by Calcein-AM/PI staining (A,C) and WST assay (B,D). Data in (B,D) are mean ± S.E. (n = 4; **P < 0.01). (E,F) Effect of H₂S on RSL3-induced lipid peroxidation. TM4 cells were exposed to 5 µM RSL3 with or without 1 mM NaHS for 16 h. After that, C11 BODIPY 581/591 staining were subjected to analysis of lipid oxidation. (G,H) Effect of H₂S on RSL3-induced ACR accumulation. TM4 cells were incubated with 10 µM RSL3 with or without 1 mM NaHS for 6 h. AcroleinRED staining was performed for ACR detection. Densitometric quantitation of relative influorescence intensity in (E,G) was performed on 20 cells randomly selected from each group, and the data were shown in (F,H), respectively (n = 20; **P < 0.01).