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. Author manuscript; available in PMC: 2017 Nov 1.

Published in final edited form as: Brain Behav Immun. 2016 Jul 28;58:291–309. doi: 10.1016/j.bbi.2016.07.158

(A) Representative image of resident microglia from the ipsilateral cortex of sham WT mice. Immunofluorescence analysis of P2Y12 (green) and CD11b (red) demonstrate colocalization of P2Y12 with CD11b in cells that have ramified morphologies indicative of surveillant microglia (inset). (B) Representative images from the ipsilateral cortex of sham and CCI WT mice at 1d and 7d post-injury. Immunofluorescence analysis of NOX2 (green), F4/80 (red) and P2Y12 (magenta) demonstrate increased NOX2 expression in F4/80+ brain macrophages at 1d post-injury, with peak NOX2+/F4/80+ expression at 7d post-injury. There was low NOX2 expression in P2Y12+ microglia at 1d post-injury, and NOX2+/P2Y12+ expression at 7d post-injury was reduced when compared to NOX2+/F4/80+ expression. Scale bar = 50μm. Inset display high magnification images from the ipsilateral cortex of sham and 7d CCI mice. (C) Representative images from the hippocampus of sham and 7d post-injury WT mice. Immunofluorescence analysis of NOX2 (green), F4/80 (red) and P2Y12 (magenta) demonstrate increased NOX2 expression in F4/80+ brain macrophages at 7d postinjury. Scale bar = 50μm. Inset display high magnification images from the ipsilateral cortex of sham and 7d CCI mice. (D) Quantification of NOX2+/F4/80+ and NOX2+/P2Y12+ colocalization after CCI in the ipsilateral cortex (CTX) and hippocampus (HP). Two-way ANOVA with Student Newman Keuls correction for multiple comparisons; data = mean ± SEM; n = 6/time point; *p<0.05, **<0.01 and ***p<0.001 versus sham group, ++p<0.01 and +++p<0.001 versus NOX2+/F4/80+ at 7d.

(A) Representative image of resident microglia from the ipsilateral cortex of sham WT mice. Immunofluorescence analysis of P2Y12 (green) and CD11b (red) demonstrate colocalization of P2Y12 with CD11b in cells that have ramified morphologies indicative of surveillant microglia (inset). (B) Representative images from the ipsilateral cortex of sham and CCI WT mice at 1d and 7d post-injury. Immunofluorescence analysis of NOX2 (green), F4/80 (red) and P2Y12 (magenta) demonstrate increased NOX2 expression in F4/80+ brain macrophages at 1d post-injury, with peak NOX2+/F4/80+ expression at 7d post-injury. There was low NOX2 expression in P2Y12+ microglia at 1d post-injury, and NOX2+/P2Y12+ expression at 7d post-injury was reduced when compared to NOX2+/F4/80+ expression. Scale bar = 50μm. Inset display high magnification images from the ipsilateral cortex of sham and 7d CCI mice. (C) Representative images from the hippocampus of sham and 7d post-injury WT mice. Immunofluorescence analysis of NOX2 (green), F4/80 (red) and P2Y12 (magenta) demonstrate increased NOX2 expression in F4/80+ brain macrophages at 7d postinjury. Scale bar = 50μm. Inset display high magnification images from the ipsilateral cortex of sham and 7d CCI mice. (D) Quantification of NOX2+/F4/80+ and NOX2+/P2Y12+ colocalization after CCI in the ipsilateral cortex (CTX) and hippocampus (HP). Two-way ANOVA with Student Newman Keuls correction for multiple comparisons; data = mean ± SEM; n = 6/time point; *p<0.05, **<0.01 and ***p<0.001 versus sham group, ++p<0.01 and +++p<0.001 versus NOX2+/F4/80+ at 7d.