Fig. 4
Validation of TRAP clients by western blot and proteomic analysis of CDG patients. a, b, d, e HeLa cells were treated with two different TRAPB-targeting siRNAs, or SEC61A1-UTR-targeting siRNA, or control siRNA, and the consequences of complex depletion were analyzed by western blots for TRAP client candidates (a), which were deduced from the proteomic analysis, possible TRAP clients (a, d), and negative control proteins (d). Where indicated, silencing phenotypes were rescued by the indicated complementation and analyzed by western blots (b, e). Quantitative western blot data represent the mean protein levels (normalized to ß-actin) relative to control and standard errors of the mean (s.e.m.). Western blots refer to at least three independent experiments. c, f In addition, TRAP-deficient fibroblasts from CDG patients with mutations in TRAPG or TRAPD genes27 were analyzed by quantitative proteomics
