Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2011 Apr 15;124(8):1301–1307. doi: 10.1242/jcs.084277

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2011.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0), which permits unrestricted non-commercial use, distribution and reproduction in any medium provided that the original work is properly cited and all further distributions of the work or adaptation are subject to the same Creative Commons License terms.

PMC Copyright notice

Fig. 2. — WRB is a 19 kDa membrane protein of the ER. (A) Identification of WRB–HA and HA–WRB. N-terminally or C-terminally HA-tagged forms of WRB (HA–WRB or WRB–HA) were expressed in HeLa cells. Proteins were separated by SDS-PAGE and analyzed by immunoblotting using the pre-immune serum (lanes 1–3), the anti-WRB serum (lanes 4–6) or anti-HA antibody (lanes 7–9). The asterisks indicate incomplete WRB. (B) Membrane association of WRB–HA. Untransfected HeLa cells (lanes 1–3) or HeLa cells transfected with HA–WRB (lanes 4–6) were analyzed by SDS-PAGE and immunoblotting using anti-WRB, anti-TRAM and anti-GAPDH antibodies. HeLa cells were permeabilized with digitonin, and cell constituents separated into a cytosolic and membrane fractions by centrifugation. Proteins of whole cell lysate (lanes 1 and 4), membrane (lanes 2 and 5) and cytosolic fractions (lanes 3 and 6) were analyzed. (C) Localization of WRB–CFP. CFP-tagged WRB (WRB–CFP) or CFP were expressed in RPE-1 cells and visualized by fluorescence microscopy. (D) ER localization of WRB–HA. WRB–HA was expressed in RPE-1 cells, immunostained with an anti-HA antibody and visualized with a fluorescent secondary antibody (red) by fluorescence microscopy. As an ER marker, calreticulin was visualized similarly using an anti-calreticulin antibody and a fluorescent secondary antibody (green).