spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.


Figure 2


Fig. 2. The µstpCys element prevents secretion of ssYFP and targets it to ERAD. (A) Schematic presentation of the Myc-tagged ssYFP vectors. (B) Hela cells expressing a vector encoding Myc-tagged ssYFP (myc), or HEK293 cell vectors encoding ssYFP-µstpCys (Cys) or ssYFP-µstpSer (Ser), were incubated with (+) or without (–) ALLN and MG-132, pulse-labeled with [35S]methionine-[35S]cysteine and chased for the indicated time with (+) or without (–) ALLN and MG-132. Cells and medium were separated, cells were lysed and substrates were immunoprecipitated (IP) from lysed cells or medium by antibodies against GFP or Myc. Immunoprecipitates were resolved by SDS-PAGE, electroblotted and blots were exposed to autoradiography ([35S]). ssYFP-µstpCys without N-glycan (*), resulting from de-glycosylation or inefficient glycosylation. ssYFP-µstpSer with high-mannose (arrow) or complex (arrowhead) N-glycans are indicated. (C) The graphs, representative of three independent experiments, illustrate the amounts of the indicated ssYFP fusion proteins remaining in untreated (open symbols) or ALLN- and MG-132-treated (+ Inhib; filled symbols) cells. The amounts detected in cells (circles) or recovered from medium (squares) were estimated by densitometry of autoradiograms in B, calculated as a percentage of their levels at the end of the pulse (100%), and the half-life values (see text) were calculated.





Right arrow Return to article