Fig. 8: In vivo bone formation was inhibited by circ_AFF4 knockdown. | Cell Death & Disease

Fig. 8: In vivo bone formation was inhibited by circ_AFF4 knockdown.

From: Circular RNA AFF4 modulates osteogenic differentiation in BM-MSCs by activating SMAD1/5 pathway through miR-135a-5p/FNDC5/Irisin axis

Fig. 8

BM-MSCs were transfected with si-circ_AFF4 or siRNA negative control (NC) and further cultured in OM for 7 days. BALB/c nude mice were injected with the BM-MSCs above and femurs were removed after 8 weeks. A ARS and ALP staining assays were performed to compare the bone cell formation in mice treated with si-circ_AFF4 or si-NC. B Representative images of H&E and Masson staining in mouse femurs showing the in vivo bone formation and collagen deposition, respectively. C Expression of the osteogenic differentiation-related transfection factor RUNX2 was evaluated using IHC staining. qRT-PCR (D) and WB (E) were conducted to determine the expression levels of osteogenic marker genes ALP, BMP4, RUNX2, Spp1, and Colla2 in femur samples from mice treated with si-circ_AFF4 or siRNA NC. The data were shown as mean ± SD based on at least three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001.

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