Fig. 2.
BMSC-EVs relieved CDFs fibrogenesis induced by TGF-β in vitro. (A) Cell proliferation ability of CDFs was determined using BrdU incorporation assay after TGF-β stimulation with or without 20 μg/ml BMSC-EVs and 50 μg/ml BMSC-EVs incubation for 24h. Green signal = BrdU. Scale bar = 180 μm. (B) The cell migration ability of CDFs was determined by wound-healing assay after 12h and 24h incubation with different treatments. (C–D) Quantification of BrdU and wound-healing data (n = 3). Data are presented as mean ± SD. *P < 0.05, ***P < 0.001, #< 0.05, ##P < 0.01, # compared with TGF-β group. (E) Cell proliferation ability of CDFs was further determined using CCK-8 assay. (n = 4) Data are presented as mean ± SD. *P < 0.05, ***P < 0.001. (F) Immunofluorescence was used to detect relative expression and distribution of α-SMA and Collagen 1 with different treatments. Green, red, and blue signal represented Collagen 1, α-SMA, and nucleus, respectively. Scale bar = 25 μm. (G) The representative image of the ability of collagen contraction of CDFs in different groups (yellow dotted line indicated the collagen gel). (H) The relative quantification of the area of collagen gel in different groups compared to that of group NC at 24h. (n = 3) Data are presented as mean ± SD. *P < 0.05, **P < 0.01. (I) Relative fluorescence intensity of Col 1 and α-SMA after different treatments. (n = 3) Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (J) Collagen 1, Vimentin and α-SMA protein levels in CDFs were presented by western blot after different treatments. (K) Quantification of western blot data (n = 4). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.