QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 3. Overexpression of AP180-C inhibits the Cu²⁺-stimulated endocytosis of PrP^C. SH-SY5Y cells stably expressing PrP^C were seeded onto coverslips and transiently transfected with either an expression plasmid encoding Myc-tagged AP180-C or an expression plasmid encoding AP180-N. After 24 hours, cells were processed as described in Fig. 2, with AP180-C detected using a polyclonal anti-Myc primary antibody and Marina Blue goat anti-rabbit IgG secondary antibody, and AP180-N detected using a polyclonal anti-AP180 primary antibody and Alexa488-conjugated donkey anti-goat IgG secondary antibody. In AP180-N-transfected cells, 3F4-labelled PrP was detected using an Alexa594-conjugated rabbit anti-mouse IgG secondary antibody. (A) Cells expressing AP180-C are unable to endocytose either transferrin or PrP^C, whereas endocytosis is unaffected in AP180-N-expressing cells. (B) Quantification of (A). Results are the mean±s.e.m. of four individual experiments, in each of which 30 cells were visualised to determine their ability to endocytose transferrin and PrP^C. The number of AP180-C- and AP180-N-transfected cells showing endocytosis is shown as a percentage of the number of untransfected cells showing endocytosis. (C) AP180-C cells in which clathrin-mediated endocytosis is blocked are not senescent, as shown by the uptake of dextran. Bars, 10 µm.

Return to article