Fig. 2. Cellular localization of (pro)insulin expressed in 3T3L1 adipocytes. (A) Colocalization of (pro)insulin and GLUT4 in the same vesicle. 3T3L1 adipocytes were infected with Adex1CA pchi at an MOI of 20 pfu/cell, then they were fixed with 2% PFA and immunostained with both rabbit polyclonal anti-GLUT4 and mouse monoclonal anti-insulin antibodies followed by respective secondary antibody treatment (GLUT4: FITC, insulin: rhodamine). Immunofluorescent staining was observed by confocal laser microscopy with band-pass filter. Bar, 5 µm. (B) (Pro)insulin and GLUT1 are differently distributed in 3T3L1 adipocytes. After cells were infected and fixed as in A, they were immunostained with both rabbit polyclonal anti-GLUT1 and monoclonal insulin antibodies as described in A. Antibody complexes were visualized with appropriate secondary antibodies coupled to rhodamine (GLUT1) and FITC (insulin). Note the different immunofluorescent staining pattern between the peripheral staining of GLUT1 (rhodamine) and vesicular distribution of (pro)insulin (FITC) in the same cell with confocal laser-microscopy.

Return to article