Supporting information for Ros-Baró et al. (October 2, 2001) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.211341698.
Fig. 12.
Nystatin reduces GLUT4 internalization on insulin removal in isolated rat adipocytes. Isolated rat adipocytes were incubated in the absence (A and D) or in the presence of 100 nM insulin for 30 min (B, C, E, and F). Thereafter, they were incubated for 15 more min with no addition (A–C) or with 50 mg/ml nystatin (D–F). Some cells (C and F) were subjected to insulin removal by washing the cells with Krebs–Ringer 10 mM 2-(N-morpholino)ethanesulfonic acid (Mes) buffer (pH 6.0) and in the absence (C) or presence (F) of nystatin, and cells were incubated in the same Krebs–Ringer Mes buffer containing 3.5% BSA and 2 mM sodium pyruvate with no addition (C) or with nystatin (F) for 15 min and processed to obtain PM lawns. After fixation, GLUT4 was immunolocalized by using a specific antibody. (Scale bar = 10 mm.)