Fig. 2: Per2 is upregulated via the MAPK signaling pathway during microglial ferroptosis.

a The differentially expressed genes (DEGs) between primary microglia treated with DMSO and 1600 μM iron + 1 μM RSL3 were identified with the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Rich factor (%) was the ratio of the number of DEGs annotated in a pathway (as indicated in the x-axis) to the number of all genes annotated in the pathway. b Volcano plot showing fold changes in genes in primary microglia treated with DMSO or 1600 μM iron + 1 μM RSL3. c qRT-PCR was performed to detect 2 ferroptosis-related genes and to verify the expression of Per2 in primary microglia treated with DMSO and 1600 μM iron + 1 μM RSL3 for 6 h (n = 3 biological repeats for each group). d Representative immunoblots of the phosphorylation status of P38 and the expression of Per2 at different time points after ferroptosis induction (the repeated immunoblots are shown in Supplementary Fig. 3c–e). e, f Quantification of the phosphorylation status of P38 and the expression of Per2 at different time points after ferroptosis induction (n = 4 biological repeats for each group). g, h Representative immunoblots (the repeated immunoblots are shown in Supplementary Fig. 3f) and quantification of the phosphorylation status of P38 after treatment with P38 MAPK inhibitor (n = 6 biological repeats for each group). i, j Representative immunoblots (the repeated immunoblots are shown in Supplementary Fig. 3g) and quantification of the expression of Per2 after treatment with P38 MAPK inhibitor (n = 6 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), one-way ANOVA with Tukey’s multiple-comparisons test (e, f, h, j). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant.