Fig. 5

Microvascular endothelial cells induce pericyte-fibroblast transition via the PDGF-BB/PDGFRβ signaling pathway in vitro. a, b Western blot analysis (a) and quantification (b) of PDGF-BB in bEnd.3 cells transfected with siNC or siRNAs targeting Pdgfb. c The expression levels of PDGF-BB in bEnd.3 cells transfected with siNC or siPdgfb#2 followed by myelin debris treatment were detected by ELISA. d, e Western blot analysis (d) and quantification (e) of PDGF-BB in bEnd.3 cells treated as described above in c. f Experimental schematic diagram of pericyte phenotypic transition transfected with siNC or siPdgfb#2 followed by myelin debris treatment. g Immunostaining of NG2 (green, upper panel), FSP1 (green, middle panel), and vimentin (green, lower panel) in primary pericytes treated as described above in f. h Quantification of the percentage of NG2⁺, FSP1⁺, and vimentin⁺ pericytes in g. i Experimental schematic diagram of pericyte phenotypic transition blocked with the PDGFRβ inhibitor imatinib (a selective PDGFRβ inhibitor) or Su16f (a specific PDGFRβ inhibitor) followed by Mye-CM. j Immunostaining of NG2 (green, upper panel), FSP1 (green, middle panel), and vimentin (green, lower panel) in primary pericytes treated as described in i. k Quantification of the percentage of NG2⁺, FSP1⁺, and vimentin⁺ pericytes in j. Scale bars: 25 μm (g and j). Data are expressed as mean ± s.e.m. n = 3 independent cultures. **p < 0.01 and ***p < 0.001 by one-way ANOVA followed by Tukey’s post hoc test in b versus siNC, and k. *p < 0.05, **p < 0.01, and ***p < 0.001 versus siNC by unpaired two-tailed Student’s t test in c, e, and h