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. 2015 Oct 9;36(6):1497–1506. doi: 10.3892/ijmm.2015.2363

Figure 2.

Figure 2

Overexpression of miR-21 enhances bone morphogenetic protein (BMP)9-induced alkaline phosphatase (ALP) activity and calcium deposition in murine multilineage cells (MMCs). miR-21 potentiates BMP9-induced ALP staining. (A) Subconfluent C2C12 cells and (B) MEFs were treated with green fluorescent protein (GFP)-conditioned medium (CM), BMP9-CM, miR.NC (100 nM) + BMP9-CM, miR-21 (100 nM) + BMP9-CM, or anti-miR-21 (100 nM) + BMP9-CM. The staining results were recorded at a lower (A and B, top panel) and higher magnification (A and B, bottom panel). ALP staining was conducted following 7 days of differentiation. miR-21 potentiates BMP9-induced matrix mineralization. (C) Subconfluent C2C12 cells and (D) MEFs were treated with GFP-CM, BMP9-CM, miR.NC (100 nM) + BMP9-CM, miR-21 (100 nM) + BMP9-CM, or anti-miR-21 (100 nM) + BMP9-CM. The Alizarin red S staining of matrix mineralization results were recorded at a lower (C and D, top panel) and higher magnification (C and D, bottom panel). Alizarin red S staining was conducted following 14 days of differentiation. Each assay condition was done in triplicate. The results were repeated in at least 2 independent batches of experiments. NC, negative control. Lower magnification refers to cells in a 24-well plate being scanned under a scanner (ScanMaker 3600). Higher magnification refers to cells being scanned under a bright field microscope (×200).