Fig. 2: TGF-β2 promoted pericyte differentiation and proliferation in vitro.

a Primary pericytes isolated from Col1α1-GFP transgenic mice were immunostained with PDGFRβ and DAPI. Green, GFP; Red, PDGFRβ; Blue, DAPI. Scale bar, 50 μm. b The mRNA expression of ACTA2 in primary pericytes with or without TGF-β2 treatment. c The protein expression of α-SMA in primary pericytes with or without TGF-β2 treatment. d Immunofluorescence analysis of α-SMA in mouse primary pericytes 2 days after TGF-β2 treatment. Green, GFP; Red, α-SMA; Blue, DAPI. Scale bar, 50 μm. e CCK8 analysis of primary pericytes treated with or without TGF-β2. f Cell counting measurements. The number of viable cells was measured by a hemocytometer. g EdU assay to assess cell proliferation. Cell nuclei with high DNA replication activity (EdU-positive cells) were stained red. Scale bar, 50 μm. h Transwell migration assay. The migrated cells were counted in three fields to determine the average number of migrated cells per field. Scale bar, 200 μm. i The protein expression levels of mTOR, Akt, Smad2, ERK and S6 were measured by Western blotting in primary pericytes treated with TGF-β2 for different times. The data are presented as the mean ± SD, and statistical analyses were performed by Welch’s t test. ns, nonsignificant difference. *P < 0.05, **P < 0.01, ***P < 0.001.