Overexpression of myc-tagged hrs-2 alters the localization of the M6PR

 

At low levels of myc-hrs-2 expression, the mannose-6-phosphate receptor (M6PR) shows a normal pattern of perinuclear, punctate localization. At higher levels of myc-hrs-2 expression, giant early endosomes form, the M6PR is no longer restricted to the perinuclear area, and a portion of the M6PR becomes localized to the surface of the induced aberrant organelles.

 

Control human fibroblasts were transfected with pDNA encoding a full length, wild type construct of myc-hrs-2. Cells were plated onto coverslips and fixed 3 days post-transfection. Cells were permeabolized with 1% saponin and coverslips were incubated with mouse-anti-myc and rabbit-anti-M6PR, followed by FITC-anti-mouse and rhodamine-anti-rabbit antibodies.