Fig. 5.

Anxa2 knockdown ameliorated immune inflammatory response and apoptosis of BV2 microglial cells induced by OGD/R.A, the mRNA expression levels of pro-inflammatory marker genes iNOS and COX2 and anti-inflammatory marker genes Arg1 and CD206 in shCtrl and shAnxa2 BV2 cells with or without OGD/R treatment. B, representative blot images and quantitative analysis of iNOS and Arg1 protein expression levels detected by Western blot in shCtrl and shAnxa2 BV2 cells with or without OGD/R treatment. GAPDH was an internal normalized control. C, the relative mRNA levels of inflammatory mediators TNF-α, IL-1β, and IL-6 in shCtrl or shAnxa2 BV2 cells after OGD/R by RT-PCR. D, the expression levels of TNF-α, IL-1β, and IL-6 in supernatants of shCtrl or shAnxa2 BV2 cells after OGD/R were measured by ELISA assay. E, cell viability accessed by CCK8 assay in shCtrl or shAnxa2 BV2 cells after OGD/R. F, cell death accessed by LDH release in shCtrl or shAnxa2 BV2 cells after OGD/R. G and H, representative blot images and quantitative analysis of apoptosis-related proteins cleaved caspase3, Bax, and Bcl2 in shCtrl or shAnxa2 BV2 cells after OGD/R detected by Western blot. All data were shown as mean ± SEM. Statistical significance was determined by two-way ANOVA for multiple comparisons. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. ∗∗∗∗p < 0.0001. All experiments were repeated at least three times. Anxa2, annexin A2; IL-6, interleukin-6; IL-1β, interleukin-1beta; iNOS, inducible nitric oxide synthase; LDH, lactate dehydrogenase; OGD/R, oxygen-glucose deprivation and reoxygenation; TNF-α, tumor necrosis factor-α.