Fig. 6

Inhibition of miR-3129-5p attenuated osteogenesis of hBMSCs. A–C Expression of miR-3129 (A), SLC7A11 mRNA and protein (B), and osteoblast markers (C) in MSCs at 48h (A, B) and day 18 (C) after transfection of miR-3129-5p and miR-3129-3p inhibitors. The levels of miRNA and mRNA expression were quantified by qPCR. The amounts of miRNA and mRNA transcripts were expressed relative to the amounts of U6sn and GAPDH transcripts, respectively. B The levels of SLC7A11 protein were detected by Western blotting. β-actin was used as a loading control. The images cropped from the data (n=1) on Fig. S4 are shown on the right side of Fig. 6B (see Additional file 7). D, E Alizarin Red S and alkaline phosphatase staining were performed in three independent experiments on day 21 after transfection of miRNA inhibitors (D), followed by quantification with ImageJ software (E). Western blotting was performed one time (n = 1 in B). Data are expressed as mean ± SD from three independent experiments (each n = 3 in A–H). *p<0.05, ***p<0.001 vs. NC inhibitor by Student’s t-test. NC, negative control; RQ, relative quantification