Fig. 3

CircRNASIPA1L1 promotes osteogenic differentiation of SCAPs. a The protein levels of ALP, RUNX2, and OSX in the sh-SIPA1L1 group and si-SIPA1L1 group as detected by Western blotting. GAPDH served as an internal control. Histograms showed the quantification of band intensities. b Transfection efficacy of circSIPA1L1. c Relative mRNA expressions of ALPL, RUNX2, and OSX measured by qRT-PCR. GAPDH was used for normalization. d Images of alkaline phosphatase (ALP) staining in the different groups after being cultured in osteogenic medium (OM) for 5 days. Histograms show ALP activity by spectrophotometry. e Alizarin Red S staining and CPC assay were conducted to investigate the mineralization of SCAPs at day 14 (scale bar = 200 μm). f ALP and RUNX2 in SCAPs transfected with si-SIPA1L1 or NC were observed by immunofluorescence staining. Scale bar = 50 μm. Quantification was done by ImageJ. *P < 0.05, **P < 0.01, and ***P < 0.001