Fig. 6: Per2 facilitates microglial ferroptosis via inhibiting Gpx4.
a The DEGs between the control and Per2OE microglia were identified with KEGG pathway enrichment analysis. b Volcano plot showing fold changes in genes in the control and Per2OE microglia. c, d The expression of Gpx4 was measured by qRT-PCR (n = 4 biological repeats for each group) and western blotting (the repeated immunoblot is shown in Supplementary Fig. 6a) after overexpressing by lentivirus. e Quantification of relative levels of Gpx4 protein (n = 6 biological repeats for each group). f, g Following a 24-h incubation with 500 nM selenium, a Gpx4 agonist, ctrl, and Per2OE microglia were exposed to DMSO or 1600 μM iron + 1 μM RSL3. Then, the cell viability was determined by CCK-8 assay (n = 3 biological repeats for each group), and the expression of ferroptosis marker Acsl4 was measured by immunofluorescence staining. Scale bar, 20 μm. h Quantification of the relative fluorescence intensity of Acsl4 (n = 3 biological repeats for each group). All data are presented as the mean ± SD. P values were determined by two-tailed unpaired Student’s t-test (c, e), one-way ANOVA with Tukey’s multiple-comparisons test (f, h). ***P < 0.01, ****P < 0.0001.
