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. 2019 May 31;38(13):e101067. doi: 10.15252/embj.2018101067

Figure 4. TGIF1 inhibits Twist1 transcriptional activity.

Figure 4

  • A
    PL45 stably expressing both Dox‐inducible TGIF1 and Tam‐inducible Twist1 (Twist1ER) were treated with Tam and/or Dox for 48 h and analyzed for Twist1 expression by immunoblotting or qRT–PCR (n = 6).
  • B
    MIAPaCa‐2 cells were transfected with wild‐type or mutated (Twist1 binding site) Twist1 luciferase reporters (TGTLuc) together with increasing amounts of Twist1 expression vector in the presence of empty vector (EV) or TGIF1 expression vector. Luciferase activity was measured 48 h following transfection and normalized on the basis of co‐transfected Renilla luciferase (n = 6).
  • C
    Total lysates from pancreas of control, KC, or KTC mice were pooled and analyzed by immunoblotting using antibodies to E‐Cadherin, Vimentin, p16Ink4A, and β‐Actin (n = 6).
  • D–F
    Expression of Cdh1, Vimentin, and p16Ink4A in pancreas from control, KC, or KTC mice (same as in C) was analyzed by qRT–PCR (n = 6).
  • G
    FFPE pancreatic sections from control, KC, or KTC mice were stained with H&E or immunostained with antibodies to E‐Cadherin, Vimentin, or p16Ink4a and revealed by IF or IHC. Representative pictures at 20× are shown (n = 30). Scale bars, 200 μM.
Data information: For (A, B, D, E, and F), data are expressed as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant; based on a two‐tailed Student's test.