Fig. 7

Circ-ITCH promotes osteogenic differentiation via recruiting IGF2BP2 in BM-MSCs under microgravity simulation. Human BM-MSCs were overexpressed circ-ITCH and silenced IGF2BP, and transfected BM-MSCs were cultured in osteogenic differentiation medium under microgravity simulation for 14 days. (A) The protein level of BRCA1 was detected by western blot. (B) The mRNA stability of BRCA1 was monitored in the presence of transcription inhibitor actinomycin D. (C) The ubiquitination of HOXC10 in BM-MSCs were assessed by Co-IP assay. (D) The interaction between BRCA1 and HOXC10 were detected by Co-IP assay. Whole lysates or normal IgG served as an input or negative control, respectively. (E) The mineralization process was monitored by ARS staining in BM-MSCs. (F) The protein levels of osteogenic differentiation markers ALP, OPN and OCN in BM-MSCs were detected by western blot. *P < 0.05, **P < 0.01, and ***P < 0.001