Fig. 3

AU-mediated osteogenic response is inhibited by noggin (BMP antagonist) and LDN193189 (type I BMP receptor inhibitor). Confirming canonical BMP signaling blockade. a, b Phospho-Smad1/5/9, BMP2, COL1A1 and RUNX2 were quantified in hBM-MSCs after 3 days of treatment with AU and/or noggin by western blot analysis and normalized to GAPDH and tSmad1. c, d Phospho-Smad1/5/9, COL1A1 and RUNX2 were quantified in hBM-MSCs after 3 days of treatment with AU and/or LDN193189 by western blot analysis and normalized to GAPDH and tSmad1. e, f, i, j Representative images and quantification of immunofluorescence staining of COL1A1 and RUNX2. Scale bars, 100 μm. g, h Representative images of ALP and ARS. k, l Quantification of ALP and ARS. Scale bars, 500 μm. All of the experiments were performed in triplicates. Data are expressed as mean ± SD. Statistical significance is denoted in the graphs. *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001