Figure 2

RT-PCR analysis and staining of INS1-miR-21 cells identify reduced expression of genes impacting β-cell function and identity. (A) The qRT-PCR analysis demonstrated that transcripts for MafA, Nkx6.1, Ins1, Ins2, Neurod1, and Glut2, were all decreased after miR-21 induction in INS1-miR-21 cell lines. (B) The qRT-PCR analysis determined that transcripts for β-cell progenitor markers Ngn3, Nanog, L-myc, and a gene associated with β-cell dysfunction in the context of dedifferentiation, Aldh1a3, were all increased after miR-21 induction in INS1-miR-21 cell lines. Western blot analysis and quantification demonstrated a trend toward a decrease in the levels of Pdx1 and significantly decreased levels of MafA (C–E). (F) Staining for Nkx6.1 showed a trend towards a decrease in the nuclear to cytoplasmic ratio in INS1-miR-21 cells as compared to controls. (G)Staining for L-Myc demonstrated a trend toward an increase in nuclear expression in INS1-miR-21 cell lines. n = 3–9 (3–4 transductions of cells); ∗p < 0.05.