Fig. 5.

ZIP8 regulates ferroptosis of monocytes under sepsis conditions in vitro.

a. Representative immunoblots of ZIP8-G, ZIP8-N, GPX4, FTH1 and xCT in RAW264.7 cells treated with different concentrations of LPS and durations. b-f. Immunoblot analyses of ZIP8-G (b, n = 4/group), ZIP8-N (c, n = 4/group), GPX4 (d, n = 4/group), FTH1 (e, n = 4/group), and xCT (f, n = 4/group) exhibited a sequential pattern of expression changes between ZIP8 (peaked at 6 h post-treatment) and ferroptosis related GPX4 and FTH1 (hit the lowest at 12 h post-treatment). g. Representative immunoblots of ZIP8-G, ZIP8-N, GPX4, FTH1 and xCT in RAW264.7 cells with/without decreased Slc39a8 expression and/or LPS treatment. h-l. Immunoblot analyses of ZIP8-G (h, n = 5/group), ZIP8-N (i, n = 5/group), GPX4 (j, n = 5/group), FTH1 (k, n = 5/group), and xCT (l, n = 5/group) exhibited that suppressing ZIP8 attenuated the decrease of GPX4, FTH1 and xCT in response to LPS. m-o. Quantification analysis showed that decreasing ZIP8 alleviated the LPS-induced upregulations of MDA (m, n = 6/group) and ferrous ions (o, n = 6/group) and reversed the decreased GSH level (n, n = 6/group). One-way ANOVA was performed in b-f. Two-way ANOVA was performed in h-o. Compared to PBS treated si-NC group: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; Compared to LPS treated si-NC group: ^#p < 0.05, ^##p < 0.01, ^####p < 0.0001.

Abbreviations: NC: normal control.