Fig. 6
In vitro degradation of p53 and MAGI-3 by HPV His6-E6 proteins. HPV16 E6, MAGI-3, and p53 proteins were in vitro translated in rabbit reticulocyte lysates. a Representative autoradiography for these 35[S]-cysteine-radiolabeled proteins following SDS-PAGE for the protein translation efficiencies (as indicated). b, c Representative SDS-PAGE for degradation of MAGI-3 (b) and p53 (c) following their mixing with purified HPV His6-E6 proteins (as indicated). The reaction mixtures were sampled after 1 h (T1) and 2 h (T2) of incubation. INPUT without purified His6-E6 protein, E6 IVT in vitro transcribed E6 protein from HPV16, as positive control (d). Quantification of MAGI-3 and p53 degradation was achieved by measuring the signal intensities of protein bands with a PhosphorImager. Data represent the mean of three independent experiments ± standard deviations. Values are presented relative to the amount of MAGI-3 or p53 in the presence of water-primed reticulocyte lysate, after 2-h incubation