Figure 4.

The ferroptosis inhibitor, Fer-1, attenuates polyphyllin VI-induced cell death by inhibiting the ferroptosis of HCC cells. The HCCLM3 and Huh7 cells were treated with or without Fer-1 for 2 h prior to treatment with PPVI for 24 h, after which different assays were performed. (A and B) Changes in the MMP of HCCLM3 and Huh7 cells were observed by MMP detection with JC-1. (C and D) GSH level detection in HCCLM3 and Huh7 cells. (E and F) MDA level detection in HCCLM3 and Huh7 cells. (G and H) Fe²⁺ levels in HCCLM3 and Huh7 cells. (I) ROS level detection in HCC cells observed using DHE. (J) Fe²⁺ level detection in HCCLM3 and Huh7 cells detected using FerroOrange fluorescent probe. (K and L) Western blot analysis of GPX4 in HCCLM3 and Huh7 cells. (M) Relative mRNA expression of GPX4 in HCC cells. ^*P<0.05, ^**P<0.01, ^***P<0.001 and ^****P<0.0001 vs. control or as indicated. NS, not significant; Con, control; HCC, hepatocellular carcinoma; PPVI, polyphyllin VI; MMP, mitochondrial membrane potential; DHE, dihydroethidium; ROS, reactive oxygen species; GPX4, glutathione peroxidase 4; Fer-1, ferrostatin-1.