Fig. 5: Promotion of circ_AFF4 on osteogenic differentiation was partially reversed by silencing of FNDC5/Irisin. | Cell Death & Disease

Fig. 5: Promotion of circ_AFF4 on osteogenic differentiation was partially reversed by silencing of FNDC5/Irisin.

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

Fig. 5

BM-MSCs were co-transfected with lentiviruses expressing circ_AFF4 and si-FNDC5 or negative control siRNA. A Relative expression of FNDC5 was detected by qRT-PCR. B Irisin concentration was determined by ELISA. C Transfected BM-MSCs were further cultured in OM or NM for 14 days. ARS and ALP staining assays were performed to analyze the osteogenic differentiation. D Transfected BM-MSCs were further cultured in OM for 14 days. RUNX2 expression was detected by immunofluorescence staining using RUNX2-specific antibodies. qRT-PCR and western blot analyses were performed to evaluate the mRNA (E) and protein (F) expression of osteogenic marker genes ALP, BMP4, RUNX2, Spp1, and Colla1 in BM-MSCs. 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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