FIGURE 4.

GADD34 localization in multiple membrane compartments. A, HeLa cells expressing GADD34-GFP were stained with MitoTracker red dye to show mitochondria (MITO). Cells were also immunostained with anti-GM130 (GOLGI) or anti-calnexin (ER) and counterstained with Alexa Fluor 647- or Alexa Fluor 350-conjugated secondary antibody respectively. Top row, individual fluorescent images (in black and white) showing GFP, mitochondria, Golgi bodies, and ER. Lower row, merged color images of GADD34-GFP (green) and MITO, Golgi bodies, or ER (red). B, (1–60)-GFP, (Δ1–60)-GFP, and (1–60,V25R)-GFP were expressed in COS7 cells, which were then stained with MitoTracker dye. GFP fluorescence (green), MitoTracker (red), and the merged images (Merge) are shown. Enlarged images of areas highlighted by boxes (Inset) are also shown. C, WT and mutant GFP-(3R-L)-GADD34 proteins (green) expressed in HeLa cells, in which the mitochondria were stained with MitoTracker dye (red). Merged images (Merge) are shown.