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. 2015 Jun 30;23(8):1349–1357. doi: 10.1038/mt.2015.101

Figure 3.

Figure 3

miR-26a directly targeted Tob1. (a) Western blot analysis of Tob1 protein expression in mesenchymal stem cells (MSC) during osteogenic induction. (b) Schematic illustration of putative miR-26a target site in mouse Tob1 3′UTR and alignment of miR-26a with wild-type (WT) and mutant (MUT) 3′UTR region showing complementary base-pairing to Tob1. MSCs were cotransfected with luciferase reporter constructs carrying WT 3′UTR or MUT 3′UTR, phRL-null (renilla plasmid) and miR-26a mimics or corresponding controls. Effects of miR-26a and corresponding controls on luciferase activity of reporter constructs were determined 48 hours post-transfection. Firefly luciferase activity values were normalized to renilla luciferase activity values. (c) RT-qPCR and Western blot analysis of Tob1 expression in MSC treated with miR-26a or corresponding control. (d) RT-qPCR analysis of Alp and Ocn mRNA levels in OVX-MSC after cotransfection of miR-26a mimics and Tob1 expression vector or empty vector. Data represent means ± SD. *P < 0.05, **P < 0.01 (n = 3 mice each group).