Figure 4

Mutant EGF receptor colocalization with endocytic recycling compartment markers. HCC827 NSCLC cell line was transiently transfected with Rab11-GFP, Rab4-GFP, EHD1-GFP or EHD3-GFP constructs. After 48 hr, cells were either left untreated (A) or treated with 10 μM monensin for 2 hr (B). Cells were fixed, permeabilized, and immunostained with anti-EGFR antibody (528) (red). Images were acquired under a confocal microscope at the medial plane. Arrows indicate mutant EGFR colocalization with GFP-tagged proteins (green). Bars represent 20 μm. (C) Colocalization coefficients for the green channel (GFP) were obtained using LSM510 Image Examiner software and normalized to the colocalization coefficient from Rab11-GFP without any treatment (none).