Alteration of dynamin 2-dependent mechanisms impairs the stimulation of CFTR endocytosis by ML141 or wiskostatin treatments.

<p>CFBE-wtCFTR cells were transfected with the corresponding siRNA to deplete dynamin 2 or caveolin 1 proteins 48 h before performing quantification of PM-CFTR or of internalized CFTR, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118943#pone.0118943.g002" target="_blank">Fig. 2</a> legend. Histograms summarizing analyzed data are displayed. Results were compared with negative control siRNA transfection conditions. (A) Dynamin 2 depletion elicited an increase of PM-CFTR amount, (B) correlated to lower CFTR internalization, (A and B) whereas caveolin 1 depletion had no impact upon PM- and internalized CFTR amounts. Then, 48 h after transfection with indicated siRNA, cells were treated with 10 μM ML141 for 30 min or 10 μM wiskostatin for 120 min, before labelling and during the 5 min internalization period. Results were compared to vehicle treatment conditions. (C) When dynamin 2 is depleted, CFTR internalization increase upon pharmacological treatments is abolished. Data represent means ± SEM of 3 independent experiments, each performed in duplicate. ***: p<0.001, **: p<0.01, *: p<0.05, ns: non significant.