Co-localization of CHC22 and components of the GLUT4 trafficking pathway in regenerating human muscle.

A, B and C) Transverse sections of skeletal muscle from a patient with PM were immunostained with a polyclonal antibody against GLUT4 (red) and monoclonal antibodies against A) embryonic myosin heavy chain (eMHC, green), B) CHC22 (green) or C) CHC17 (green). Regenerating myofibers were identified by centrally located nuclei stained with DAPI (blue in merge). Boxed regions in the merged images are magnified seven-fold at the right showing overlap of markers in yellow (scale bars, 20 µm). D and E) Transverse sections of skeletal muscle from a patient with PM were immunostained with a polyclonal antibody against CHC22 (red) and a monoclonal antibody against VAMP2 (green). Regenerating myofibers were identified by centrally located nuclei stained with DAPI (blue in merge). Boxed regions in the merged images are magnified seven-fold at the right showing intracellular co-localization (yellow) of CHC22 and VAMP2 in a regenerating fiber of D) small diameter or E) large diameter (scale bars, 20 µm). At the far right, the percent internal staining for the markers indicated on the y-axis was quantified as in Figure 1F, for fibers that were characterized according to internal staining for the marker indicated on the x-axis. n is the number of fibers analyzed in each staining combination shown at the left. Significant increased internal staining was determined by Student’s t-test (*p<0.05, **p<0.01).