Fig. 7.

FSP1 protects hepatocytes against ferroptosis by attenuating mitochondrial iron loading and mtROS production. (A) OVX hepatocytes were treated for 24 h by 100 μM FAC in the presence or not of 10 μM iFSP1, and then stained by PI. (B) The effects of stable expression of V5-FSP1 on iron metabolism and antioxidant defense genes in HepG2 cells. (C) HepG2 cells with the stable expression of V5-FSP1 or the backbone control (pLX304) were treated with or not 100 μM FAC for 24 h, followed by PI staining. (D) HepG2/V5-FSP1 and control cells were treated for 4 h with 200 nM RSL3 prior to PI staining. The percentages of PI⁺ cells (A, C and D) were quantified, and the data shown are mean ± SEM (n = 9 fields of 3 independent experiments). (E) HepG2/V5-FSP1 and control cells were treated for 4 h with 200 nM RSL3, followed by staining for mitochondrial Fe²⁺ with 5 μM Mito-FerroGreen (MFG) and mtROS with 5 μM MitoSOX. The fluorescence intensity of MFG staining was quantified and is presented as mean ± SEM (n = 30 cells of 3 independent experiments). *, P < 0.01. Bars, 100 μm.