Fig. 6
Validation of physicochemical properties of TRAP clients. a–h Plasmid driven over-production of model precursor polypeptides in HeLa cells was carried out in the presence of the indicated siRNA for 24 h. Where indicated, MG 132 was present during the last 8 h. Samples were analyzed by western blot. The identity of the mature proteins is based on experiments where N-glycosylation was inhibited by Tunicamycin (Supplementary Fig. 6a-h). We note that the pre-form represents the precursor polypeptide, the form without any addition the mature protein without N-glycosylation, g the mono-gylcosylated mature protein, and gg the doubly-glycosylated mature protein. i In case of the experiments with MG 132, quantitative western blot data represent the mean precursor and mature protein (N-glycosylated plus non-glycosylated) levels plus the corresponding dot plots for at least three independent experiments. The western blots for the depleted proteins are shown in Supplementary Fig. 6i. *, unspecific band
