Fig. 7. Suppression of PIKfyve alters the steady-state distribution of the CI-MPR and enhances receptor degradation. (A) HeLa cells were treated for 72 hours with control siRNA, PIKfyve-specific siRNA duplex II or a combination of siRNA duplexes II and V. Cells were fixed and stained against CI-MPR and EEA1 prior to imaging. Suppression of PIKfyve results in a redistribution of the CI-MPR from the TGN to peripheral punctae that show considerable overlap with the EEA1-labelled compartment. Boxed areas are magnified in right corner to show degree of overlap. Bars, 20 µm. (B) HeLa cells were treated for 72 hours with either control, PIKfyve-specific siRNA duplex II or a combination of PIKfyve-specific siRNA duplexes II and V. Cells were lysed and lysates blotted for PIKfyve, EGF receptor, transferrin and CI-MPR, and tubulin (loading control). Representative results are shown. (C) Quantification of data from four independent experiments.