Fig 2.
Specific cleavage of PABP by the EMCV 3C proteinase in uninfected cells. HEK293 cells were transiently transfected with mammalian expression vectors encoding EMCV WT 3C protease (WT 3Cpro), a catalytically inactive mutant form of 3C (H46A C159A), or EGFP. Forty-eight hours after transfection, total protein was isolated, fractionated by SDS-PAGE, and analyzed by immunblotting with the indicated antibodies. Full-length PABP runs around 70 kDa, whereas the cleavage (clvg) fragment migrates faster, around 45 kDa. Transfection efficiency indicated by EGFP fluorescence was above 60%. An asterisk indicates a cross-reacting band representing a cellular protein nonspecifically recognized by the polyclonal PABP antibody. The migration of molecular mass standards (in kilodaltons) is indicated to the left of the panel.