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. Author manuscript; available in PMC: 2015 Jan 10.
Published in final edited form as: FASEB J. 2006 Feb 2;20(6):705–707. doi: 10.1096/fj.05-4661fje

Figure 3.

Figure 3

Activation of Ins-R/PI3K/Akt pathway occurs 2 min after insulin stimulation and requires caveolin-3 expression. Control (A, B, C) and caveolin-3 null (B, C) skeletal muscle cells were differentiated for 5 days and treated with or without insulin (160 nM) for 2, 5, and 20 min. Cell lysates were immunoprecipitated with anti-insulin receptor-β IgGs (A) and IRS-1 IgGs (B), and immunoprecipitates were subjected to immunoblot analysis using specific antibody probes against phospho-tyrosine proteins (A), and p85 (B). In addition, total cell lysates were separated by SDS-PAGE and subjected to Western blotting analysis using anti-phospho-Akt and Akt IgGs (C). Total IRS-1 and p85 protein expression was not affected by insulin (data not shown). Moreover, total cell lysates from control and caveolin-3 null myotubes were subjected to immunoblot analysis with anti-Ins-Rβ IgGs (D). Finally, control and caveolin-3 null myotubes were left untreated or treated with 10 μM pervanadate for 15 min, and cell lysates subjected to immunoblot analysis using anti-phospho-Akt and Akt IgGs (E).