Figure 5.

Chronic stress induced microglial activation in mPFC. (A) Experimental protocol for FACS was presented. (B) Gate strategy for CD11b⁺45^Low microglia was presented. (C) Concentration of IL-1β in mPFC was measured (two-way ANOVA with Tukey’s post hoc testpost hoc, ***P < 0.001, compared with CONT; n = 6). (D) Representative flow cytometry images showing the percentage of CD11b⁺CD45^LowMHC-II⁺ microglia in mPFC of CONT, CUMS, and CORT groups, as well as the statistical analysis (two-way ANOVA with Tukey’s post hoc testpost hoc, ***P < 0.001, compared with CONT; n = 6) were presented. (E) Representative histogram images showing CD11b⁺CD45^Low microglial ROS in mPFC were presented. (F) Representative histogram images showing CD11b⁺CD45^Low microglial Clec2d on the membrane and intracellular in mPFC were presented. (G) Representative histogram images showing CD11b⁺CD45^Low microglial intracellular TLR9 in mPFC were presented. (H) Representative Western blots showing microglial Clec2d, TLR9, NF-κB pathway, and NLRP3 inflammasome were presented. Relative expression of Clec2d, TLR9, p-p65, p-IκBα, NLRP3, ASC, pro-caspase-1, cleaved caspase-1, pro-IL-1β, and cleaved IL-1β was analyzed (two-way ANOVA with Tukey’s post hoc testpost hoc, *P < 0.05, **P < 0.01,***P < 0.001, compared with CONT; n = 3).