Fig. 6: Silencing of STK25 decreases phenotypic switching in human aortic smooth muscle cells.

hASMCs were transfected with STK25 siRNA or NTC siRNA. a Representative images of cells processed for immunofluorescence with anti-Ki67 (pink) antibody; nuclei stained with DAPI (blue). Quantification of the staining. b Proliferation rate was assessed by measuring the DNA synthesis using a Biotrack cell proliferation ELISA. c Representative images of migrated cells stained with crystal violet after 6, 12, and 24 h. Quantification of the staining. d Representative images of calcium deposition. Quantification of the staining. e, f Relative mRNA e and protein f abundance of selected calcification markers. Protein levels were analyzed by densitometry; representative Western blots are shown with vinculin used as a loading control. In (a, c, d), the scale bars represent 25 and 50 µm, respectively. Data in (a–e) are mean ± SEM from 5 to 8 wells per group; data in (f) are mean ± SEM from 11 to 12 wells per group. OA, oleic acid. *P < 0.05, **P < 0.01, ***P < 0.001 for STK25 siRNA vs. NTC siRNA.