Fig. 1: Primary Acsl4 knockout hepatocytes are resistant to ferroptosis induction.
A Quantification of cell viability by trypan blue staining of Hepa1-6 cells after 24 or 48 h treatment with DMSO as untreated control, 3 µM RSL3, 200 nM Liproxstatin-1 (Lipr-1) or both. B–G Primary hepatocytes were isolated from 8 to 12-week-old mice. B Quantification of cell viability of Acsl4f/f hepatocytes by trypan blue staining after 24 h treatment with DMSO as untreated control, 1 µM RSL3, 1 µM erastin, 5 µM FIN56, 10 µM FINO2, or 1000 µM arachidonic acid (AA). C Quantification of cell viability by trypan blue staining of Acsl4f/f hepatocytes after 24 or 48 h treatment with DMSO as untreated control, 3 µM RSL3, 200 nM Lipr-1, or both. D Acsl4f/f hepatocytes were treated for 24 h with DMSO as untreated control, 3 μM RSL3, 200 nM Lipr-1, or both, and stained with BODIPY 665/676 for 1 h. Lipid peroxidation was determined as mean fluorescence intensity (MFI) by flow cytometry. E TUNEL staining of Acsl4f/f hepatocytes after 24 or 48 h treatment with DMSO as untreated control, 3 µM RSL3, 200 nM Lipr-1, or both. F Quantification of percentage of TUNEL-positive from all (DAPI-positive) cells. G Quantification of cell viability by trypan blue staining of Acsl4∆hepa hepatocytes after 24 or 48 h treatment with DMSO as untreated control, 3 µM RSL3, 200 nM Lipr-1, or both. Data are expressed as mean ± SEM from one representative out of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test comparing each group with untreated cells for each time point.
