Loss of function of LRP5 resulted in damaged osteogenesis in vitro and in vivo.

<p>(A) The relative absorbance of MC3T3-E1 cells transfected with a serial dilution of siLRP5 in the WST-1-based colorimetric assay. (B) Representative 3D images of bone microstructure in the knee-joints in LRP5 WT and KO mice. (C) Quantification of trabecular bone volume fraction (BV/TV) in the knee-joints in WT or KO mice was determined from the μCT measurement. (D) Relative expression levels of SMAD7 in the bone tissues of mice. * p < 0.05, ** p < 0.01, n = 3/group.</p

The expression dynamics of effectors of miR-375-3p during osteogenesis.

<p>(A) Relative expression levels of LRP5 during MC3T3-E1 differentiation and osteogenesis. (B) Relative expression levels of β-catenin during MC3T3-E1 differentiation. (C) Relative expression levels of β-catenin in bone tissues from osteoporotic patients and their controls. * p < 0.05. ** p < 0.01.</p

The examples of region of interests.

<p>The examples of region of interests.</p

miR-375-3p damaged osteogenesis by inducing cell apoptosis.

<p>(A) Relative expression levels of RUNX2 in MC3T3-E1 cells treated with miR-375-3p mimics. (B) Relative expression levels of SOST in MC3T3-E1 cells treated with miR-375-3p mimics. (C) Representative TUNEL staining images of MC3T3-E1 cells. More TUNEL positive staining cells (Red) were found after the cells were transfected with miR-375-3p mimics. (D) Quantification of TUNEL-positive cells. * p < 0.05, ** p < 0.01. n = 3/group.</p

miR-375-3p targeted LRP5 and β-catenin.

<p>(A) The binding sites between miR-375-3p and the 3’UTR of LRP5 and β-catenin. (B) The relative luciferase activity of MC3T3-E1 cells transfected with luciferase-wild type (or mutant) LRP5 plasmids and miR-375-3p mimics. (C) The relative luciferase activity of MC3T3-E1 cells transfected with luciferase-wild type (or mutant) β-catenin plasmids and miR-375-3p mimics. * p < 0.05. ** p < 0.01, NS, not significant. n = 3/group.</p

miR-375-3p arrested the expression of LRP5 and β-catenin.

<p>(A) Relative expression levels of LRP5 in MC3T3-E1 cells upon transfection of miR-375-3p mimics or inhibitors. (B) Relative expression levels of β-catenin in MC3T3-E1 cells upon transfection of miR-375-3p mimics or inhibitors. (C) Representative staining images of MC3T3-E1 cells. Less green cells were found after the cells were transfected with miR-375-3p mimics. * p < 0.05. ** p < 0.01. n = 3/group.</p

Sequences of all primers.

<p>Sequences of all primers.</p

The baseline of mice.

<p>The baseline of mice.</p