Figure 6.

IRS2 promotes membrane localization of GLUTs during decidualization. HESC were treated for 72 hours with DC following siRNA-mediated knockdown of IRS2. Membrane-associated GLUT expression was observed using Western blot (A) and immunofluorescence (B). (A) Membrane associated and cytoplasmic protein fractions were isolated as described in Materials and Methods. These fractions were subjected to Western blot and probed with antibodies for GLUT1 and GLUT4. Integrin β3 and HSP90 were used as fraction-specific controls. Densitometry of membrane fraction is provided (right). (B) Immunofluorescent staining using antibodies specific for GLUT1 and GLUT4. DAPI was used as a nuclear stain and phalloidin was used to stain actin filaments.