Figure 7

Colocalization of AmpA with calnexin, golvesin and p25. A) SomeAmpA is found in a perinuclear compartment and colocalizes there withthe ER marker calnexin. Top panels are Wt control cells that do notcontain the calnexin GFP or the AmpA tap tag fusion protein. The cellswere fixed, permeabilized and stained with rabbit anti-TAP antibody andgoat anti-rabbit antibody conjugated to FITC (red). The second rowpanels show AmpA tap tag cells containing calnexin GFP (green) fixed,permeabilized and stained with rabbit anti-TAP antibody and goatanti-rabbit antibody conjugated to FITC (red). Cells were incubated withDAPI to stain the nuclei (Blue). Arrows indicate areas of calnexin andAmpA tap tag colocalization in a perinuclear compartment. Scale bars are18 um for the field of cells in top row and 9 um for thezoomed images in the second row. Images are single optical sections froma confocal z-series. B) AmpA is found in the Golgi colocalizingwith N-golvesin. mRFP-AmpA cells were transformed with a plasmidcarrying N-Golvesin-GFP (Golgi marker-green) incubated, fixed andstained with rat anti-RFP antibody followed by Alexa goat anti-ratantibody (red). Arrow indicates the area of colocalization in the Golgi.DAPI (blue) stains the nucleus. Images are single optical sections froma z-series. Scale bar is 11um. C) AmpA colocalizes with p25, arecycling endosome marker in a perinuclear compartment. mRFP-AmpA cellswere stained with rat anti-RFP (green) and mouse anti-p25 (red). DAPI(blue) stains the nucleus. Arrow indicates area of p25 and AmpAcolocalization. Images are single optical sections from a z-series.Scale bar is 11 um.