Dysregulation of the Core-Clock Genes (BMAL1, PER2, and NR1D1) and SMAD4 Leads to Differential Cell Proliferation and Apoptosis in Panc1 and AsPC1 Cells
(A) Proliferation analyses of PDA cell lines for KD conditions (shBMAL1, shPER2, shNR1D1, and shSMAD4), SMAD4-OEs, and corresponding empty vectors (shCtrl and oeCtrl, respectively).
(B) Apoptosis analysis of PDA cell lines after KDs or OEs. Data are expressed as mean ± SEM, n ≥ 6 (A and B).
(C–F) Representative images of apoptosis assays for Panc1 shCtrl and knockdown cells shBMAL1, shPER2, shNR1D1, shSMAD4 (C) or Panc1 oeCtrl and SMAD4-OE cells (D) and AsPC1 shCtrl and knockdown cells shBMAL1, shPER2, shNR1D1 (E) or AsPC1 oeCtrl and SMAD4-OE cells (F), with fluorescently labeled caspase3/7 with (objective 20×) at time points 0, 72, and 120 h, respectively. Data are expressed as mean ± SEM, n ≥ 6 (A and B).