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Fig. 3 DHE inhibited the activation of NF-κB signaling and the NLRP3 inflammasome in LPS-treated BV2 microglial cells via TREM2 activation. Cells were pretreated with DHE for 1 h and then exposed to LPS for another 24 h. (A–D) The levels of total/phosphorylated IKKα/β (A), IκB α (B), and P65 (C) were measured by Western blot. (D–E) The expression levels of total NF-κB p65 in the cytoplasmic fraction (D) and nuclear fraction (E) were quantified by Western blot. (F) The nuclear translocation of p65 was evaluated by immunofluorescence analysis. Cells were stained with anti-p65 antibody (green) and DAPI (blue). (G) Cells were transfected with negative control shRNA or TREM2 shRNA, and then the NF-κB p65 expression level in BV2 cells after treatment with DHE or LPS was assessed by Western blot analysis. (H) The mRNA levels of NLRP3 and IL-1β were detected by RT-PCR analysis. (I–L) Protein levels of NLRP3 (I), ASC (J), cleaved caspase-1 (K), and pro-IL-1β (L) were measured by Western blot analysis. (M) Cells were transfected with TREM2 shRNA or a negative control shRNA, and after being treated with DHE or LPS, the level of NLRP3 expression in BV2 cells was determined by Western blot analysis (∗∗p < 0.01 and ∗∗∗p < 0.001 by the one-way ANOVA). Data are presented as the mean ± SD analyzed by one-way ANOVA. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 vs. the LPS-treated group; #p < 0.05, ##p < 0.01, and ###p < 0.001 vs. the control group; N.S. = not significant; n = 3.