Fig. 2: Ferroptosis regulates SIO-induced human primary HRPEpiC cell death.

Fer-1 (50 µM) or its vehicle (0.14% DMSO) was added to human primary HRPEpiC cells for 3 h before control (PBS) or SIO (10 mM) treatment. DFO (25 µM) or its vehicle (PBS) was added to human primary HREpiC cells 3 h before control (PBS) or SIO (10 mM) treatment. A Phase-contrast and fluorescence microscope images of cell morphology and Live/dead cell staining of SIO-exposed human primary HRPEpiC cells with/without Fer-1 pre-treatment are shown. Scale bar = 100 µm. B CCK-8 cell viability assay results of the effect of Fer-1 on 10 mM SIO-induced human primary HRPEpiC cell death. n = 4 (independent experiments). C CCK-8 cell viability assay results of the effect of DFO on 10 mM SIO-induced human primary HRPEpiC cell death. n = 4 (independent experiments). D Phase-contrast and fluorescence microscope images of cell morphology and Live/dead cell staining of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment are shown. Scale bar = 100 µm. n = 4 (independent experiments). E Labile iron levels in SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment. n = 4 independent experiments. F Total GSH level of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment. n = 4 (independent experiments). G GSSG level of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment. n = 4 (independent experiments). H GSH level of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment. n = 4 (independent experiments). I GSH/GSSG ratio of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment. n = 4 (independent experiments). J The expression of GPx-4 of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment showed by Western blot. K Quantitative analysis of the expression of GPx-4. n = 4 (independent experiments). L The expression of ACSL-4 of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment showed by Western blot. M Quantitative analysis of the expression of ACSL-4. n = 3 (independent experiments). N MDA level of SIO-exposed human primary HRPEpiC cells with/without Fer-1 pre-treatment. n = 4 (independent experiments). O MDA level of SIO-exposed human primary HRPEpiC cells with/without DFO pre-treatment. n = 4 (independent experiments). *p < 0.05, **p < 0.01, ***p < 0.001.