Fig. 5.

PKCβII, GLUT4 and MARCKS bound actin in an insulin-dependent manner. L6 myotubes were treated for 30 min with and without 10 μIU/ml insulin, treated with cross-linking agent and then immunoprecipitated (IP) with anti-actin antibody. The bound proteins were then extracted by magnetic separation and treated with dithiothreitol to break cross-links. a Proteins were separated by SDS-PAGE and subjected to immunoblotting (IB) for actin, PKCβII, GLUT4, MARCKS and the insulin receptor. b–f Densitometric analysis (as described in the legend of Fig. 1) for the relevant proteins in (a) are shown beneath the blots. The experiments were repeated on two occasions in separate experiments with similar results. Ins, insulin; cyt, cytosol fraction; memb, membrane fraction