(A) Upper panel: Glutathione sepharose-bound GST-C2 pro D137A was prephosphorylated with [γ-32P]ATP and incubated in extracts ± G6P or Bcl-xL. Samples of sepharose-bound proteins were analyzed by autoradiography. Lower panel: In parallel, extracts treated with the same conditions were analyzed for C3 activity using the caspase substrate Ac-DEVD-pNA. n = 3
(B) Radiolabeled in vitro translated full-length C2 WT or D137 was incubated in extracts ± G6P. Samples were retrieved over time and analyzed by SDS-PAGE and autoradiography.
(C) Glutathione sepharose-bound GST-C2 pro D137A was prephosphorylated with [γ-32P]ATP and incubated in buffer with His-PP1 that had been pre-incubated in extract. Samples of sepharose-bound proteins were analyzed by autoradiography. n = 2
(D) Percent survival of Xenopus oocytes microinjected with full-length C2 WT or D137A mRNA. n = 3