Fig. 6.

Ubiquitination assay of CFBE-wt cells overexpressing Nedd4–2. A–C: IP was conducted with either an anti-CFTR antibody (M3A7) or a control antibody (IgG). WB were probed for Nedd4–2 (A), ubiquitin (FK2 antibody, B), or wt-CFTR (C-term antibody, C). Control and Nedd4–2 samples were run on the same gel, but are separated for presentation. D: summary of Nedd4–2 levels in control and Nedd4–2 experiments (N = 3). E: summary of the effects of Nedd4–2 overexpression on ubiquitinated wt-CFTR, which is reported as the amount of ubiquitinated wt-CFTR (FK2) divided by the amount of total wt-CFTR determined by Western blot (N = 3). ***P < 0.001.