Bone marrow mesenchymal stem cells induce M2 microglia polarization through PDGF-AA/MANF signaling

Figure 4 Mesencephalic astrocyte–derived neurotrophic factor expression in microglia/macrophages in cerebral ischemia. A and B: Western blot (A) and qRT-PCR analysis (B) of MANF expression at 24 h after cerebral I/R injury; C: Immunohistochemical analysis of MANF expression in the microglia/macrophages of the injured brains. Representative images of immunohistochemical staining for the microglia/macrophage marker Iba1 (green) and MANF (red), with DAPI (blue) as a nuclear counterstain, are shown. ^aP < 0.05, ^bP < 0.01 vs Sham group; ^dP < 0.01 vs I/R and I/R + PBS groups; ^fP < 0.01 vs I/R + BMSCs and I/R + BMSCs-NC groups. Scale bar = 50 μm. Arrows point to the MANF⁺/Iba1⁺ cells. The values are expressed as the mean ± SD (n = 6). MANF: Mesencephalic astrocyte–derived neurotrophic factor; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; I/R: Ischemia/reperfusion; PBS: Phosphate-buffered saline; BMSCs: Bone marrow mesenchymal stem cells; BMSCs-NC: Negative control-transfected BMSCs; BMSCs-siMANF: MANF siRNA-transfected BMSCs; Iba1: Ionized calcium-binding adapter molecule; DAPI: 4'6-diamidino-2-phenylindole.