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Fig. 4. The actin-disrupting agent cytochalasin D affects GLUT4myc translocation differently in response to insulin and PDGF. Serum-deprived GLUT4myc myoblasts were treated without (a-c) or with (d-f) 250 nM cytochalasin D for 20 minutes before and during incubation with either 50 ng ml–1 PDGF-BB or 100 nM insulin for 7 minutes at 37°C. The cells were then either (A) fixed, permeabilized and stained for filamentous actin with rhodamine-phalloidin or (B) left intact and reacted with an anti-Myc antibody and subjected to the colorimetric assay for cell surface GLUT4myc. The increase in surface GLUT4myc in response to each stimulus over basal (unstimulated) conditions was calculated and presented as the proportion of the response in the absence of cytochalasin D. The means±s.e.m. are shown of four independently performed experiments, each performed in triplicate. *, P<0.05 compared with the respective control in the absence of the drug. Scale bar, 10 µm.
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