(A) Western blot analysis for Twist in WT and KRasG12D PDEC cultures at day 8. α-Tubulin serves as a loading control.
(B) Quantitative RT-PCR analysis of Twist in WT and KRasG12D PDEC cultures at day 0 and day 8. Error bars indicate SD (n = 3).
(C) Quantitative RT-PCR analysis of Twist in KRasG12D PDEC cultures 8 days after infection with recombinant lentiviruses encoding shRNA targeted against GFP (control shRNA) or Twist (Twist shRNA3 or Twist shRNA5). Error bars indicate SD (n = 3).
(D) Western blot analysis for Twist and senescence effectors in KRasG12D PDEC cultures 8 days after infection with recombinant lentiviruses encoding control shRNA, Twist shRNA3, or Twist shRNA5. Equal loading was verified with anti-α-Tubulin.
(E) Quantitative RT-PCR analysis of p16INK4A in KRasG12D PDEC cultures 8 days after infection with recombinant lentiviruses encoding control shRNA, Twist shRNA3, or Twist shRNA5. Error bars indicate SD (n = 3). *, p-value < 0.05.
(F) SA-β-gal staining of KRasG12D PDEC cultures 8 days after infection with recombinant lentiviruses encoding control shRNA or Twist shRNA3. Scale bar: 100 µm.
(G) Quantification of SA-β-gal staining in KRasG12D PDEC cultures 8 days after infection with recombinant lentiviruses encoding control shRNA, Twist shRNA3, or Twist shRNA5. Error bars indicate SD (n = 6 FOV). *, p-value < 0.05. Data are representative of three independent experiments. See also Figure S2.