Figure 4.

A) Effect of miR-342 mimic on IA-2β. A) IA-2β protein level 72 h after transfection with the miR-342 mimic in MIN6 cells. A representative Western blot of the endogenous IA-2β and α-tubulin, with protein extracts from MIN6 cells 72 h after transfection with a control sequence or miR-342 mimic (left). Average of adjusted density values after Image J quantification on 3 different blots normalized to α-tubulin (right). Experiments were repeated 3 times, and similar results were obtained. Error bar, sd. *P < 0.015. B) RIP PCR analysis, with anti-Ago2 antibody or anti-IgG (background control) used to assess the changes in enrichment of miRNA:mRNA in silencing complexes after miR-342 over expression for 48–72 h in MIN6 cells with Ptprn-, Ptprn-, and Gad2-specific primers, relative to the input fraction (10% of material before immunoprecipitation). Enrichment was quantified with qReal-time PCR. Fold change is presented as bound fraction over input fraction, normalized to Gapdh or β-actin. Error bars, sd. Experiments were repeated at least 3 times.