Figure 5.

MG-specific Nrf2 knockdown modulates inflammatory and anti-inflammatory phenotypes of MG after ischemic stroke. Male and female Cx3cr1^CreERT2/+ (C/+), Nrf2^fl/fl-Cx3cr1^CreERT2/+ (cKD), and Nrf2^-/- (tKO) mice were subjected to MCAO. (A) On day 1 post-injury, the contralateral (C) and ipsilateral (I) hemispheres of MCAO mice were harvested, followed by mononuclear cell isolation. The isolated cells were stained with CD45 and CD11b antibodies followed by intracellular staining with CD68 antibody to determine CD45^intCD11b⁺ MG positive for CD68 expression (n=8/group). (B) The isolated cells were ex vivo cultured in the presence of Golgi Plug for 4.5 h followed by surface staining with CD45 and CD11b antibodies. Following fixation and permeabilization, cells were stained with IL-1β antibody to assess the intracellular IL-1β expression in CD45^intCD11b⁺ MG by flow cytometry (n=8/group). (C) On day 2 post-injury, the isolated cells were stained with CD45 and CD11b antibodies followed by intracellular staining with CD206 antibody to assess CD206-expressing CD45^intCD11b⁺ MG (n=8/group). The representative flow cytometry figures are shown in Supplementary Figure 2 . *p<0.05; **p<0.01; ***p<0.001; N.S.: no significant difference by two-way ANOVA.