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Figure 3
INS1-miR-21 cells directly target the Tgfb2 pathway. (A) The qRT-PCR analysis demonstrated that transcripts for Tgfb2 and Smad2 were decreased after miR-21 induction in INS1-miR-21 cells. (B–C) Western blot analysis showed decreased Tgfb2 and Smad2 protein expression in miR-21 induced INS1-miR-21 cells. (D)Tgfb2 and Smad2 mRNAs were enriched in the miR-21 biotinylated pulldown samples normalized to cell lysate and compared to C-elegans control miR-67 biotinylated mimic pulldown. (E–F): Luciferase assays showed that activity was decreased after miR-21 induction in cells expressing wild type 3′UTR, but not in cells expressing the mutated 3′UTR for (E)Tgfb2 and (F) Smad2. n = 3–6; ∗p < 0.05.