Figure 1

Incorporation of PKH26-labeled MPs into cultured HUVEC and the in vitro effect of MPs on endothelial-to-mesenchymal transition. Immunofluorescence microscopy revealed that red PKH26-labeled MPs were readily incorporated into cultured HUVEC, as shown by DAPI staining (A), white arrows). Light microscopy analysis shows the morphologic changes of HUVEC treated with TGF-β1 alone or with TGF-β1 and KMSC-derived MPs simultaneously, compared to non-treated control cells (‘Control’) (B). Western blot analysis shows CD31 and α-SMA protein expression levels in HUVEC treated with vehicle control or KMSC-derived MPs. (C). A bar graph shows the quantification of CD31 and α-SMA protein expression levels normalized by β-actin protein expression measured using the Image J program (D). Non-treated control was expressed as ‘Control’. Vehicle control of MPs comprised MP-free supernatants after centrifugation was expressed as ‘Vehicle control.’ Results are expressed as the mean ± SE of three different experiments. The Kruskal-Wallis test was performed; * P <0.05 versus non-treated control. # P <0.05 versus TGF-β1 alone. DAPI, 4',6-diamidino-2-phenylindole; HUVEC, human umbilical vein endothelial cells; KMSC, kidney-derived mesenchymal stem cells; MPs, microparticles; SE, standard error; TGF-β1, transforming growth factor- β1; α-SMA, α-smooth muscle actin.