Fig 1: EZH2 is necessary during CIS escape.a LS174T cells have been stimulated with sn38 and emergence was evaluated in the presence or not of GSK343 or DZNepA as indicated after 10 days. The quantification of emergence is presented (n = 5; *p < 0.5; **p < 0.01, ***p < 0.001). b, c LS174T cells have been transfected with siRNAs directed against EZH2 after 4 days of treatment. EZH2 expression has been detected by western blot (n = 4) and emergence was evaluated after 10 days (n = 13; ****p < 0.0001). d, e LS174T cells have been transfected with siRNAs directed against SUZ12 after 4 days of treatment. SUZ12 expression has been detected by western blot (n = 4) and emergence was evaluated after 10 days (n = 3). f PLC clones were generated as described above, emergent cells were then trypsinized and further treated with GSK343 (5 µM) and/or Palbociclib (0.5 µM) for 2 days. The percentage of senescent cells was evaluated as the number of cells expressing SA-ß-gal activity (n = 5). Proliferation of PLC was evaluated by flow cytometry using an antibody directed against the Ki67 antigen (n = 5). DNA DAPI staining was used to evaluate the percentage of cells in SubG1 phase by flow cytometry (n = 5)