AmpA is found in vesicles and in the perinuclear region but not withF-actin. A) Wt control cells (top row) and cells carrying themRFP-AmpA fusion protein construct (bottom row) were immunostained withrat anti-RFP antibody and goat anti-rat antibody conjugated to AlexaFluor 488 (Invitrogen). The left panel is anti-RFP antibody staining(green), and the right panel shows overlays of the immunostaining onDAPI stained images (blue) to mark the nucleus and transmitted images.The scale bar is 9 um. B) Western blots of cellfractionations of mRFP-AmpA expressing cells. MW is molecular weightstandard. CY-Cytoplasm, Mem-Membrane, SN-Soluble Nuclear, CB-ChromatinBound, CS-Cytoskeletal. Controls for this fractionation are in . C) AmpA does not colocalize with F-actin. Cells werefixed and stained for AmpA Tap tag and F-actin. Green: Alexa fluor 488phalloidin stain for F-actin; Red: AmpA Tap tag antibody. Representativeimages are single optical sections from a Z series. Scale bar is 5 um.An AmpA-Tap-Wt (top) and an AmpA-Tap-overexpressing strain (bottom) areshown.