Fig. 2

SIRT5 is upregulated after cerebral I/R injury. A qPCR assay indicating Sirt5 mRNA expression in primary cultured microglia at the indicated time points following OGD. n = 3 per time point. B, C Primary cultured microglia were subjected to OGD and reoxygenation at the indicated time points. Western blot assay (B) and quantification analysis (C) showing SIRT5 protein expression. n = 3 per time point. D qPCR assay indicating Sirt5 mRNA expression in cerebral tissues at the indicated time points following cerebral ischaemia injury. n = 5 per time point. E, F Mouse brain homogenates were extracted after MCAO surgery for 1 h and reperfusion at the indicated time points. Western blot assay (E) and quantification analysis (F) showing SIRT5 protein expression; n = 5 per time point. G Primary cultured microglia were subjected to OGD and reoxygenation at the indicated time points. Immunofluorescence assays of SIRT5 (green) with colabelled 4′,6-diamidino-2-phenylindole (DAPI; blue, nuclei). Scale bars, 20 μm. H The fluorescence intensity of SIRT5 was quantified using ImageJ software. Data are presented as the mean ± SEM. ns: no significance, **P < 0.01, ***P < 0.001, and ****P < 0.0001