Fig. 2.

S897A mutation abolishes ligand-independent suppression of anoikis by EphA2 in HeLa cells. (A) At 3 days after transfection, HeLa cells were detached from tissue culture plates and cultured in suspension for 24 h. Then they were collected, and apoptotic cell death was analyzed by staining of nuclei with Hoechst 33258. Examples of scored viable (upper panel) and apoptotic (lower panel) HeLa cells are shown. (B) Lysates from HeLa cells transiently transfected with control luciferase (shControl) or EphA2 shRNA (shEphA2) were analyzed by immunoblotting with antibodies against EphA2 and α-tubulin. (C) Apoptotic cells were scored as a percentage of the total number of transfected cells, and data are the means ± SEM from four independent experiments (*, P < 0.05; t-test). In one experiment, at least 100 cells were counted in randomly selected fields.