In vivo miR-215 inhibition reduces α-SMA and fibronectin expression.

<p>(A) Design of prevention study. Db/db mice (8 weeks of age) were treated with antagomir-control or antagomir-215 (80 mg /kg body weight, three injections in three consecutive days) via the tail vein, and the kidneys were harvested three weeks after the last injection. miR-215 expression was determined by qRT-PCR (B) and subsequently analyzed for CTNNBIP1 mRNA (C) and protein (D) expression levels in glomeruli from antagomir-215-treated mice by qRT-PCR and immunohistochemical staining, respectively. qRT-PCR results were normalized to U6 snRNA. Data are shown as means ± SE (n = 6 per group) (*<i>P</i><0.05 compared with antagomir-control treated mice). (E) Quantification of renal glomerular CTNNBIP1 positive staining area shown in (D). (F) Western blot analyses and quantitation of the activated and total β-catenin protein levels. Data are shown as means ± SE (n = 6 per group) (<i>*P</i><0.05 compared with antagomir-control treated mice). qRT-PCR was used to analyze α-SMA (G) and FN (H) mRNA expression levels in glomeruli of the antagomir-215-treated mice. qRT-PCR results were normalized to GAPDH. Data are shown as means ± SE (n = 6 per group) (*<i>P</i><0.05 compared with antagomir-control treated mice). α-SMA (I) and FN (J) protein expression levels were detected in the glomeruli by immunohistochemical staining and western blotting, respectively. control, db/db mice treated with antagomir-control, antagomir-215, db/db mice treated with antagomir-215. Magnification, ×400 in H.