Fig. 6: miR-320-3p inhibits the osteoblast activity and matrix mineralization of MC3T3-E1 cells by targeting Hoxa10.
From: Bone-targeted lncRNA OGRU alleviates unloading-induced bone loss via miR-320-3p/Hoxa10 axis
a qRT-PCR analysis of miR-320-3p levels during osteoblast differentiation of MC3T3-E1 cells (n = 3). b Western blot analysis of Hoxa10 protein expression during osteoblast differentiation of MC3T3-E1 cells (n = 3). c qRT-PCR analysis of Hoxa10 mRNA expression levels and d western blot analysis of Hoxa10 protein expression levels after treatment with mimic-320, inhibitor-320, or the corresponding controls (n = 3). e The relative luciferase activity of luciferase reporters containing the wild-type (WT) Hoxa10 3′UTR or the Hoxa10 3′UTR with a mutated (MUT) miR-320-binding site in 293T cells cotransfected with mimic-320 or its negative control was measured (n = 3). MC3T3-E1 cells were cotransfected with inhibitor-320 and si-Hoxa10 or its negative controls. qRT-PCR analysis of Alp, Osx, Runx2, and Ocn mRNA levels (f), western blot analysis of Osx, Runx2, and Ocn protein levels (g), relative Alp activity (h), representative images of Alp staining in MC3T3-E1 cells (i), and representative images of alizarin red staining in MC3T3-E1 cells are presented, and the relative areas of mineralization were quantified by Image J (j) (n = 3). All data are the mean ± SD. *P < 0.05, **P < 0.01.
