Fig. 3

Effect of HCG18 and miR-30a-5p on osteogenic differentiation of BMSCs. a Putative binding sites for HCG18 and miR-30a-5p. b Left panel, expression of recombinant adenovirus transfected with HCG18. Right panel, luciferase activity of HCG18-wt and HCG18-mut in HEK-293T cells treated with miR-665 mimic or NC. c RNA pull-down assay indicated that HCG18 could be enriched by biotin-labelled miR-30a-5p but not biotin-labelled miR-30a-5p mutant type. d HCG18 overexpression inhibited miR-30a-5p expression in OP-BMSCs. e ALP activity assay and f ALP staining and Alizarin Red S staining were performed on OP-BMSCs to investigated the effects of HCG18 and miR-30a-5p co-transfection on the osteogenic differentiation of OP-BMSCs. g Determination of the expression of specific markers for osteogenic differentiation (Runx2, OCN, OPN) of OP-BMSCs co-transfected with HCG18 and miR-30a-5p. h Knockdown of HCG18 promoted miR-30a-5p expression in OP-BMSCs. i ALP activity assay and j ALP staining and Alizarin Red S staining were performed on OP-BMSCs to investigate the effects of shHCG18 and miR-30a-5p-inhibitor co-transfection on the osteogenic differentiation of OP-BMSCs. k Determination of the expression of specific markers for osteogenic differentiation (Runx2, OCN, OPN) of OP-BMSCs co-transfected with shHCG18 and miR-30a-5p-inhibitor. * P < 0.05. The OP-BMSCs were from one human subject and the experiments were performed in three replicates