Fig. 4
EMPA ameliorates inflammation through the regulation of mitochondria homeostasis in microglia. A Volcano plots showing upregulated and downregulated IR-DEGs (left) and EMPA-DEGs (right) of microglia. Red and blue dots indicate upregulated and downregulated DEGs in the corresponding comparison groups, respectively. B Violin plots showing the expression of indicated genes among three groups in microglia. C Bar plots showing the GO terms related to inflammation enriched for the upregulated (red) and downregulated EMPA-DEGs (blue) of microglia. The color from light to dark indicates the statistical significance value from low to high. D Heatmap showing the expression of genes mapped to GO pathways negative regulation of NLRP3 inflammasome complex assembly from three groups in microglia. The color key from blue to red indicates the expression level from low to high. E On 3 days post-IR injury, Western blotting analysis was performed to detect the expression of S1009a9 and NLRP3 inflammasome pathway-related proteins among 3 groups. β-Actin served as the internal protein control. F–K Relative densitometry quantitation of relative protein expression levels (n = 3–4). L Bar plots showing the GO terms related to mitochondria function enriched for the upregulated (red) and downregulated EMPA-DEGs (blue) of microglia. The color from light to dark indicates the statistical significance value from low to high. M Heatmap showing the expression of genes mapped to GO pathways from three groups in microglia. The color key from blue to red indicates the expression level from low to high. N. Venn diagrams showing the intersection of genes enriched in upregulated GO terms related to mitochondria function including (a) mitophagy, (b) mitochondrial genome maintenance, (c) positive regulation of mitochondrion organization after EMPA treatment in microglia. O Western blotting analysis showed Mfn1 and Opa1 expression changes at different time points after IR injury. P On 3 days post-IR injury, Western blotting analysis was performed to detect the expression of Mfn1 and Opa1 among 3 groups. β-Actin served as the internal protein control. Q, R Relative densitometry quantitation of Mfn1 and Opa1 expression levels (n = 6)