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ERRATUM

View larger version (46K): [in this window] [in a new window] [Download PPT slide]   FIGURE 2. PonA-inducible expression of F-ARMER. A. Two HT1080 cell lines inducibly expressing F-ARMER were independently derived (clone 1 and clone 2). Cells were grown in the absence (-) or presence (+) of ponA for 3 days before processing for immunofluorescence using mouse anti-Flag antibody followed by goat anti-mouse FITC. Nuclei were stained with Hoescht stain and visualized. B. The two independently derived inducible cell lines were grown as above and analyzed by Western blot. A cell line inducibly expressing LacZ was also analyzed and used as a negative control for the ponA inducible system.

View larger version (24K): [in this window] [in a new window] [Download PPT slide]   FIGURE 5. ARMER is an ER integral membrane protein. A. Cells expressing F-ARMER were homogenized (H) and separated into membrane (M) and supernatant (S) fractions. The membrane fraction was treated with TX-114 and separated into pellet (P), lower (L), and upper (U) phases. The membrane fraction was also treated with sodium carbonate and separated into pellet (P) and supernatant (S) fractions. Proteins were isolated from each fraction and separated by SDS-PAGE for Western blot. F-ARMER fractionates as an integral membrane protein. B. ARMER co-localizes with the ER marker calreticulin. Cells expressing Flag-tagged ARMER were examined by co-immunofluorescence using a mouse anti-Flag antibody followed by goat anti-mouse FITC to detect ARMER expression, visualized in green (left panel), and rabbit anti-calreticulin followed by goat anti-rabbit Alexa 594 to detect calreticulin, an ER marker, in red (middle panels). The merged image shows extensive overlap in yellow (right panel).

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