Fig. 4

Endothelial PDGF-BB induces pericyte-fibroblast transition and extracellular matrix deposition in vitro. a The PDGF-BB expression levels in bEnd.3 cells treated with 1 mg/ml myelin debris at the indicated time points were measured by ELISA. b, c Western blot analysis (b) and quantification (c) of PDGF-BB in bEnd.3 cells treated as described above in a. d The PDGF-BB expression levels in bEnd.3 cells treated with the indicated concentrations of myelin debris for 72 h were measured by ELISA. e, f Western blot analysis (e) and quantification (f) of PDGF-BB in bEnd.3 cells treated as described above in d. g Experimental schematic diagram of pericyte phenotypic transition induced by culture medium (empty, control), EC-CM treated with PBS, Mye-CM treated with myelin debris, and PDGF-BB. h Immunostaining of PDGFRβ (red), pericyte marker NG2 (green, upper panel), and fibroblast markers FSP1 (green, middle panel) and vimentin (green, lower panel) in primary pericytes treated as in g. i Quantification of the percentage of NG2⁺, FSP1⁺, and vimentin⁺ pericytes. j, k Immunostaining and quantification of extracellular matrix collagen I (green, upper panel) and laminin (green, lower panel) in primary pericytes treated as described in g. The nuclei are stained blue with DAPI. l, m Western blot analysis (l) and quantification (m) of NG2, FSP1, and vimentin in primary pericytes treated as described above. n, o Western blot analysis (n) and quantification (o) of extracellular matrix collagen I and laminin in primary pericytes treated as described above. Scale bar: 25 μm (h and j). Data are expressed as mean ± s.e.m. n = 3 independent cultures. *p < 0.05, **p < 0.01, and ***p < 0.001 versus 0 h, 0 mg/ml or control by one-way ANOVA followed by Tukey’s post hoc test in a, c, d, f, i, k, m, and o