Fig. 2

miR-204-5p overexpression inhibits the expression of RUNX2, ALP and OCN. hADSCs were transfected with miR-204-5p mimics, miR-204-5p inhibitors or NC and cultured in PM or OM. a Relative mRNA levels of miR-204-5p, RUNX2, ALP and OCN measured by qRT-PCR on day 14 of osteogenic induction. U6 and GAPDH were used as internal control genes. The data were normalized to the NC-PM group. b Left panel: Western blot of RUNX2, ALP and OCN protein levels on day 14 after osteogenic induction. GAPDH was used as an internal control. Right panel: quantitative analysis of the protein levels. c Confocal microscopy of cells with OCN with DAPI counterstaining on day 14 after osteogenic induction. Scale bars, 20 μm. Data are presented as the mean ± SD. *hADSCs transfected with miR-204-5p mimics or inhibitors vs NC in PM; ^#hADSCs transfected with miR-204-5p mimics or inhibitors vs NC in OM. * or ^#p < 0.05, ** or ^##p < 0.01, *** or ^###p < 0.001 (n = 3 independent experiments)