Fig. 5: Microglial ferroptosis is regulated by Per2 in vitro.
a The expression of Per2 was measured by western blotting after overexpressing by RNA lentivirus. b Quantification of relative levels of Per2 protein (n = 3 biological repeats for each group). c, f Ctrl or Per2OE microglia and WT or Per2GKO microglia were treated with DMSO or 1600 μM iron + 1 μM RSL3 for 24 h, and the cell viability was determined by CCK-8 assay (n = 6 biological repeats for each group). d Immunofluorescence image showing expression of ferroptosis marker Acsl4 in ctrl or Per2OE microglia after treatment with DMSO or 1600 μM iron + 1 μM RSL3 for 18 h. Scale bar, 20 μm. e Quantification of the relative fluorescence intensity of Acsl4 (n = 3 biological repeats for each group). g Immunofluorescence image showing expression of ferroptosis marker Acsl4 in WT or Per2GKO microglia after treatment with DMSO or 1600 μM iron + 1 μM RSL3 for 18 h. Scale bar, 20 μm. h Quantification of the relative fluorescence intensity of Acsl4 (n = 4–5 biological repeats for each group). i The levels of IGF-1 in the microglia culture medium (n = 5 biological repeats for each group) were detected by ELISA kits. All data are presented as the mean ± SD. P values were determined by two-tailed unpaired Student’s t-test (b, i), one-way ANOVA with Tukey’s multiple-comparisons test (c, e, f, h). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
