Fig. 6

KCNMA1-AS1 promotes osteogenic differentiation of HBMSCs through miR-1303/COCH axis. A The predicted binding site between the miR-1303 and KCNMA1-AS1, and the luciferase activity of the WT KCNMA1-AS1 and Mut KCNMA1-AS1 in 293T cells treated with mimics miR-1303 or mimics NC. B Transfection efficacy after KCNMA1-AS1 interference is determined by qRT-PCR. C–G Western blotting (C) and quantitative analysis of COCH (D), RUNX2 (E), Osterix (F), and COL1A1 (G) in HBMSCs treated with si-KCNMA1-AS1 and si-KCNMA1-AS1 plus si-miR-1303. H, I Staining of calcium deposition by Alizarin Red (H) and quantitative analysis of staining areas (I) in HBMSCs treated with si-KCNMA1-AS1 and si-KCNMA1-AS1 plus si-miR-1303. Scale bars, 200 μm. J, K Staining of ALP (J) and quantitative analysis of staining areas (K) in HBMSCs treated with si-miR-1303 and si-miR-1303 plus si-COCH. Scale bars, 200 μm. L, M Immunofluorescence staining of RUNX2 (l) and quantitative analysis of RUNX2-positive nuclei (M) in HBMSCs treated with si-miR-1303 and si-miR-1303 plus si-COCH. Scale bars, 100 μm. N, O Immunofluorescence staining of Osterix (N) and quantitative analysis of RUNX2-positive nuclei (O) in HBMSCs treated with si-miR-1303 and si-miR-1303 plus si-COCH. Scale bars, 100 μm. A Values are shown as mean ± SD. ***P < 0.001, two-way ANOVA. B, D–G, I, K, M, O Values are shown as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, one-way ANOVA