Figure 6.

Lats2 and ASPP1 promote apoptosis of polyploid cells. (A) WI-38 cells were infected with H-RasV12 or empty retroviral vector. Two days after infection, cells were transfected with siRNA oligonucleotides specific for ASPP1 or Lats2 or control siRNA (siASPP1, siLats2, and siCont, respectively). Three days after infection, cells were harvested, fixed, stained with PI, and subjected to FACS-based DNA content analysis. (B) Graphic representation of averaged values corresponding to the indicated cell subpopulations in A. (C) HCT116 cells were transiently transfected with GFP-tagged Lats2, together with the indicated combinations of H-RasV12 and Flag-tagged Yap1. Twenty-four hours later, cells were fixed and subjected to analysis by light microscopy. Nuclear DNA was detected by DAPI staining. Single representative cells are magnified and presented in the far right panel. (D) HCT116 cells were transiently transfected with V5-tagged ASPP1, myc-tagged Lats2, and Flag-tagged Yap1. Twenty-four hours later, cells were fixed and immunostained using anti-V5 or anti-Flag antibodies. Nuclear DNA was detected by DAPI staining. Cells marked 1 and 2 are representative examples of nuclear and cytoplasmic ASPP1 staining, respectively.