Fig. 1

SENP6 is upregulated in microglia after cerebral ischaemia–reperfusion injury. A Primary microglia were challenged with OGD for 1 h and then reperfusion for 3 h, 6 h, 12 h, 24 h and 48 h. RT-qPCR confirmed the mRNA levels of Senp6. B The protein levels of SENP6 in primary microglia subjected to OGD treatment and reperfusion at the indicated times. C Quantitative analysis of the protein levels of SENP6 in B. D Density gradient centrifugation collected microglia around the ischaemic penumbra from mice for 3 h, 6 h, 12 h, 24 h and 72 h after MCAO surgery. RT–qPCR detected mRNA levels of Senp6. E The protein expression analysis of collected microglia by western blot assay. F Quantification of the protein levels of SENP6 in E. G Representative double immunostaining of SENP6 (red) with Iba-1 (a microglial marker, green), GFAP (an astrocyte glial marker, green) and NeuN (a neuronal marker, green) from ischaemic penumbra of brain tissue after MCAO surgery. H Quantification of Iba-1⁺/SENP6⁺, NeuN⁺/SENP6⁺, and GFAP⁺/SENP6⁺ fluorescence intensity was quantified using ImageJ. Scale bars, 40 µm. Data are presented as the mean ± S.E.M. from at least three dependent experiments and analysed by one-way ANOVA followed by Dunnett’s post-hoc test (A, C, D, F) or unpaired Student's t test (H). ns for P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001