Figure 5

Localization of p36 and Cb5 proteins with modified TMDs in BY-2 cells. (A) Schematic illustrations of various p36-myc and myc-Cb5 proteins with modified TMDs and their corresponding subcellular localizations in transformed (via biolistic bombardment) BY-2 cells. The name of each mutant construct denotes p36-myc or myc-Cb5 and the specific alterations to its TMD(s). Hydrophilic portions of the p36 ORF are colorized blue, with TMD1 and TMD2 in p36 colored black and white, respectively. The single TMD in Cb5 is colorized red. Grey boxes denote the position of the myc epitope. Cyt, cytosol; ER, endoplasmic reticulum; Mito, mitochondria; ?, unknown subcellular compartment(s). (B) Representative (immuno)epifluorescence micrographs illustrating the localizations of the various constructs shown in (A). Each micrograph is labelled at the top left with the name of either the transiently-expressed protein, the endogenous mitochondrial marker protein, E1β, the endogenous peroxisomal marker protein, catalase, or ConA (fluor-conjugated Concanavalin A) serving as stain for the ER [89, 98]. Hatched boxes represent the portion of the cells shown at higher magnification in the panels to the right. Solid arrowheads indicate obvious colocalizations; open arrowheads indicate obvious non-colocalizations. Bar = 10 μm.