Fig. 5

Knockdown of miR-204-5p enhanced osteogenic differentiation of SCAPs. a Western blot assay showed lower protein levels of ALP, RUNX2, and OSX in the inhibitor-NC group than the miR-204-5p inhibitor group. GAPDH was the internal control. b Grayscale analyses. c qRT-PCR showed higher levels of ALPL, RUNX2, and OSX in the miR-204-5p inhibitor group. d ALP staining and ALP activity assay were performed to evaluate the expression of ALP in the inhibitor-NC and miR-204-5p inhibitor group at day 5. e Alizarin red staining showed that the miR-204-5p inhibitor group generated more calcified nodules than the control group on day 14. CPC assay showed quantification of Alizarin red staining by spectrophotometry. f Immunofluorescence staining showed upregulated ALP and RUNX2 in the miR-204-5p inhibitor group compared with the NC group (scale bar = 50 μm). Quantification was done by ImageJ. *P < 0.05, **P < 0.01, and ***P < 0.001