CHC22 is localized at the ERGIC in HeLa GLUT4 and human myotubes. (A and B) Representative SIM of a HeLa-GLUT4 cell (HeLa-G4; A) and the human skeletal muscle cell line hSkMC-AB1190-GLUT4 (hSkMC; B) stained for CHC22 (red) and p115 (blue). The gray circles delineate the nuclei (N). Muscle cell staining with each antibody is shown in black on white below the color images. Scale bars: 10 µm. (C and D) Representative SIM of the perinuclear region of HeLa-GLUT4 cells and hSkMC-AB1190-GLUT4 stained for CHC22 (red), p115 (C), and ERGIC-53 (blue; D). The solid gray lines delineate the nuclear border (N). The dashed white lines span the segment for which fluorescence intensities for GLUT4 (green), CHC22 (red), and p115 or ERGIC-53 (blue) were plotted. Arrowheads indicate areas of peak overlap. Scale bars: 1 µm. In A–D, GLUT4 (green) was detected by GFP tag in HeLa-GLUT4 or immunostained with anti-GFP antibody in hSkMC-AB1190-GLUT4. Merged images show red/green overlap in yellow, red/blue overlap in magenta, green/blue overlap in cyan, and red/green/blue overlap in white.