Fig. 2

Knockdown of circ_AFF4 suppresses BM-MSC osteogenic differentiation

(A) BM-MSCs were transfected with shcirc_AFF4_1–4# or shNC and the expression level of circ_AFF4 in shcirc_AFF4 or shNC transfected BM-MSCs was detected using RT-qPCR. BM-MSCs that were transfected with shcirc_AFF4 or shNC were cultured in OM or NM for 14 days. Afterward, different assays were performed to assess the osteogenic differentiation. (B) Representative images of ARS and ALP staining. Scar bar = 100 μm. (C) Representative RUNX2 immunofluorescence staining image. Scar bar = 100 μm. Gene and protein expression of osteogenic markers, ALP, OPN, RUNX2 and Colla1 were detected by (D) RT-qPCR and (E) western blot. * p < 0.05, ** p < 0.001 and *** p < 0.001. Each experiment was performed at least three times independently