FIGURE 6.

DLT regulated microglial phagocytic receptor TLR2/4 through 5HT_2AR/cAMP/PKA/CREB/GR signaling. (a) Immunofluorescence assay and (b) its quantification results demonstrated that treatment of DLT or AAV‐si‐5HT_2AR stimulated GR nuclear translocation in hippocampal microglia of APP/PS1 mice (brain slice, n = 4; cell, n = 12). Scale bar: 10 µm. (c, d) RT‐PCR results demonstrated that co‐treatment of GR inhibitor Mifeprex abolished the upregulation of TLR2/4 induced by the treatment of DLT or si‐5HT_2AR in BV2 cells (n = 3). (e) Immunofluorescence assay and (f) its quantification results demonstrated that co‐treatment of CREB inhibitor 666‐15 abolished the promotion of GR nuclear translocation induced by the treatment of DLT or si‐5HT_2AR in GR‐U2OS cells (n = 4). Scale bar: 100 µm. (g, h) Results of transactivation and mammalian one‐hybrid assays demonstrated that co‐treatment of 666‐15 abolished the promotion of GR transactivation induced by the treatment of DLT or si‐5HT_2AR in HEK‐293 T cells, implying that the DLT‐mediated GR activation was independent of the direct combination of GR with DLT (n = 3). All values were presented as the mean ± SEM. For animal tissue assays, ^## p < 0.001 compared with WT group by t test. *p < 0.05, **p < 0.01, ***p < 0.001 compared with APP/PS1 group mice by two‐way ANOVA. For cell assays, *p < 0.05, **p < 0.01, ***p < 0.001 compared with si‐Ctrl by one‐way ANOVA