Fig. 7

SIRT5 promotes OGD/R-induced expression of proinflammatory cytokines and chemokines in microglia. A Primary microglial cells were transduced with recombinant adenovirus carrying the SIRT5 coding sequence or the shRNA sequence. qPCR assay indicating the mRNA expression levels of Il-1β, Il-6, Tnf-α, Cxcl1 and Ccl2 in primary cultured microglia from the indicated groups. B ELISA analysis showing the protein secretion levels of IL-β, IL-6, TNF-α, Cxcl1 and Ccl2 in primary cultured microglial cell supernatants from the indicated groups. C, D Western blot assay (C) and quantification analysis (D) indicating the protein expression levels of iNOS and CD16/32 in primary cultured microglia. E Immunofluorescence assays of iNOS (green) and Iba-1 (red) with colabelled DAPI (blue, nuclei). Scale bars, 20 μm. F The fluorescence intensity of iNOS and Iba-1 was quantified using ImageJ software. Data are presented as the mean ± SEM from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001