Figure 4

Macrophage colony-stimulating factor (M-CSF) upregulates antidepressant signaling pathways and brain-derived neurotrophic factor (BDNF) expression. (a) Western blot analysis of BDNF, extracellular signal-regulated kinase (ERK)1/2, Akt, and glycogen synthase kinase-3β (GSK3β) in the prefrontal cortex (PFC, ctx), cerebellum (ceb), and hippocampus (hip) of the wild-type (WT) and knockout (KO) mice. (b) Western blot analysis of BDNF, ERK1/2, Akt, GSK3β, synaptophysin, PSD95, VGLUT1, GFAP, and Iba-1 in the synaptosomes isolated from the PFC of the WT and KO mice. (c) Western blot analysis of ERK1/2, Akt, GSK3β, synaptophysin, PSD95, VGLUT1, GFAP and Iba-1 in the synaptosomes isolated from PFC slices of the WT mice after a time-dependent M-CSF stimulation (10 nM). (d) Western blot analysis of BDNF, ERK1/2, Akt, and GSK3β in BV2 and HT22 cells from the co-culture set. (e) Western blot analysis of BDNF, ERK1/2, Akt, and GSK3β in HT22 cells after M-CSF stimulation (40 ng/ml) for 24 h. (f) Western blot analysis of ERK1/2, Akt, and GSK3β in HT22 cells after a time-dependent M-CSF (40 ng/ml) stimulation. (g) Western blot analysis of BDNF, ERK1/2, Akt, and GSK3β in HT22 cells treated with RAW264.7 cultured medium with or without STAT3 inhibition. Each quantification of western blotting was obtained with relative densitometry and normalized with α-tubulin. Whole western blotting images of BDNF are presented in Supplementary Figure S11. Data are means±SEM and *p<0.05, **p<0.01, and ***p<0.001. Representative data from three independent experiments are shown.

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