AS160 regulates Rab10 recruitment by insulin stimulation. (A) Adipocytes were transfected with EGFP-Rab10 and either mCherry-AS160-4P or mCherry-AS160-4P, R/A. Insulin-stimulated Rab10 vesicle recruitment to the cell periphery was followed using TIRF microscopy. AS160 images were taken using the epifluorescence mode. (B) EGFP-Rab10-QL and GLUT4-mCherry were transfected into adipocytes, and dual-color TIRF microscopy images were taken before and 6 min after insulin stimulation. Bars, 4 µm. (C) Rab10 vesicle density quantification. Rab10 vesicle densities were measured before and 6 min after insulin perfusion, and all densities were normalized to the mean of those measured from control cells before insulin stimulation. Data are represented as mean ± SEM (error bars). Control, n = 9 cells; AS160-4P, n = 10 cells; AS160-4P, R/A, n = 9 cells; Rab10-QL, n = 12 cells. **, P < 0.02; *, P < 0.05.