Fig. 5

PPARγ activation mediates effects of GRb1-treated microglia in vitro. (A) Representative micrographs after immunostaining against Iba1 display microglial activation phenotype. Fluorescence intensity reflects PPARγ expression. Scale bars: 20 μm. (B) Quantification of expression of PPARγ in mRNA levels. (C) Quantitation of PPARγ fluorescence intensity. (D) Representative western blot of of PPARγ and p-PPARγ protein expression. (E) Quantification of PPARγ and p-PPARγ protein (n = 5). (F) The unbiased stereological quantification of microglial cell area, and (G) total length of processes. (H) Experimental protocol to test the effects of conditioned medium from microglial cultures on NPCs. (I) NPC cultures were maintained in native medium (NM) and directly stimulated by GRb1, LPS, and GW9662; or they were maintained in conditioned medium (CM) of activated microglia. After 2 h or 10 days, the proliferation and differentiation of NPCs were tested, respectively, by immunolabeling BrdU (upper two rows) or DCX and GFAP (lower two rows). Scale bars: 15 μm. (J, K) The ratio of BrdU⁺-DAPI⁺ to DAPI⁺ cells in NPC cultures maintained in native medium or exposed to conditioned medium from microglial cultures. (L, M) The ratio of DCX⁺-DAPI⁺ to DAPI⁺ cells in NPC cultures maintained in native medium or exposed to conditioned medium. (N, O) Relative mRNA levels of DCX in treated groups (n = 6). The statistical results are shown in Table S8. *P < 0.05, ** P < 0.01, *** P < 0.001