figs3.

Validation of Pax6 overexpression and Pax4 silencing in INS-1E cells. (A) Significantly decreased Scgn mRNA expression was observed 48 h after transfection of INS-1E cells with a vector carrying shScgn. In contrast, transfection of INS-1E cells with CMV-Scgn (48 h) significantly increased Scgn mRNA content. (B–B₁) Western blotting confirmed successful Scgn silencing and overexpression by shScgn and CMV-Scgn, respectively. Total protein load (Cy5) was used to normalize protein expression. Representative immunoblot is shown. Tbp mRNAs were used to normalize gene expression. Data were expressed as means ± s.d. from triplicate experiments, ***p < 0.001, *p < 0.05 (one-way ANOVA).3