Fig. 7

Macrophage Caspase 6 signaling-mediated ASK1 activation regulates hepatocyte ferroptosis during inflammatory action. BMMs were transfected with the p-CRISPR-ASK1 KO or control vector followed by LPS stimulation; (A) Western blotting and relative intensity of HMGB1 in LPS-stimulated BMMs, N = 6/group. B ELISA analysis of HMGB1 secretion in culture medium, N = 6/group. C Western blotting analysis of HMGB1 in ASK1 activated BMMs isolated from Caspase 6.^KO mice after LPS stimulation, N = 6/group. D HMGB1 release from hepatocytes, N = 6/group. E Schematic figure for macrophage/hepatocyte co-culture system; (F-G) LDH release and MDA concentration of hepatocytes in co-cultures, N = 5/group. H Western-assisted analysis and relative intensity of NOX1 and GPX4 in hepatocytes after co-culture, N = 6/group. I BMMs from Caspase 6-KO mice were transfected with the p-CRISPR-ASK1 Act or control vector followed by LPS stimulation. MDA concentration in hepatocytes was detected, N = 6/group. Ferroptosis-related proteins were measured in hepatocytes after co-culture by using western blot assay, N = 6/group (J) and Immunofluorescence staining, N = 4/group (K). All data represent the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001