Figure 7.

Effect of miR-10b on LEC marker K17 expression in HCEC and in normal human organ-cultured corneas by immunostaining and western blot. (a) Transfection with miR-10b increased, whereas with miR-10b inhibitor decreased staining for K17 in HCEC compared to their corresponding controls. (b) MiR-10b treatment in human organ-cultured corneas led to increased expression of K17 compared to scrambled mimic control or Inhibitor. The same exposure time was used for each set of compared immunostained sections, and the assessment was done by more than one observer. The pictures are representative of two to three independent experiments. (c) Western blot analysis, total extracted protein from transfected HCEC with miR-10b mimic (M) or its inhibitor (I) and their corresponding controls, mimic control (MC) and inhibitor control (IC), respectively, was separated on gradient SDS-PAGE gels, transferred to membrane and probed with K17 antibody (Table 2). Antibody to β-actin was used as loading controls and for semi-quantitation. miR-10b treatment decreased, whereas its inhibitor increased, protein level of K17. (d) Quantitation of K17 protein level. The bar graph represents average ± SEM of pooled values (n = 4) of densitometric scans. *P < 0.05, compared with scrambled control values by paired two-tailed t test.