Fig 5. Fas death domain pY is important for non-death functions of Fas.

(A) SW480 cells were transiently transfected with control or Fas siRNA before BrdU incorporation analysis by using microplate-based method. Means ± SEM of three independent experiments are shown (*** p < 0.001, unpaired t test). Typical reduction of Fas level, based on immunoblot, is shown in the inset. (B) SW480 cells stably expressing control (LacZ) or indicated Fas proteins carrying silent mutations at the site targeted by an siRNA against Fas were transiently transfected with control or Fas siRNA for 48 h. They were subsequently synchronized to G1 phase by serum deprivation for 24 h and then treated with 1 ng/ml of sFasL for 30 min before analysing the increase in proliferation by BrdU incorporation measurement using microplate-based method. Means ± SEM of three independent experiments are shown (* p < 0.05, ** p < 0.01, *** p < 0.001, unpaired t test). (C) Fluorescently prelabeled SW480 cells were subjected to Boyden chamber migration assay using FluoroBlok membrane inserts. Cells were untreated or induced to migrate by 1 ng/ml sFasL for 2 h. Cells that migrated through the membrane were fixed, imaged, and counted. Data were presented as percentage of cells that migrated through the membrane compared to control cells (LacZ). Means ± SEM of three independent experiments are shown (* p < 0.05, ** p < 0.01, unpaired t test). Numerical values underlying the data summary displayed in this figure can be found in S1 Data.