Fig. 5.

ABCC5 negatively regulates ferroptosis by stabilizing SLC7A11. (A) Western blot analysis determined the expression of SLC7A11 in HuH7-SR cells. (B,C) Western blot analysis detected the expression of ABCC5 and SLC7A11 in HuH7-SR cells after knockdown and overexpression of ABCC5. (D,E) HuH7-SR cells were treated with ML385 (5 μM), brusatol (50 nM),MK571 (10 μM), Erastin (10 μM) and RSL3 (3 μM) for 24 h, and the protein expression of ABCC5 and SLC7A11 in HuH7-SR cell lines were determined by qRT-PCR and Western blot assays. (F) The ABCC5-knockdown HuH7-SR cells were treated with sorafenib (10 µM) with or without Erastin(10 μM) for 24 h, and the levels of glutathione (GSH) and lipid ROS were assayed (n = 3, * P < 0.05 relative to the control group). (G) Western blot assays detected the protein expression of GPx4 in HuH7-SR cells . (H) HuH7-SR cells were treated with sorafenib (10 µM) with or without inhibitors (LY294002, 10 µM; Brusatol, 50 nM; MK571, 10 µM; Erastin, 10 µM; RSL3 3 µM) for 24 h. Western blot evaluated the protein expression of p-AKT, AKT, NRF2, ABCC5, SLC7A11 and GPx4. *P < 0.05, ** P < 0.01, # P > 0.05.