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. Author manuscript; available in PMC: 2015 May 22.
Published in final edited form as: Cell Rep. 2014 May 10;7(4):1227–1238. doi: 10.1016/j.celrep.2014.04.006

Figure 7. Adiponectin Stimulates the Interaction between APPL1 and IRS1.

Figure 7

(A) Serum-starved C2C12 myotubes were pre-treated with or without adiponectin (1 μg/ml, 10 min) and then with or without insulin (10 nM, 3 min). Endogenous APPL1 was immunoprecipitated. Coimmunoprecipitated IRβ or IRS1 and phosphor-ylated IR, IRS1, Akt, GSK3β, or APPL1 and their protein levels in cell lysates were detected by western blot analysis with specific antibodies as indicated.

(B) APPL1-suppressed C2C12 myocytes were transfected with myc-tagged and RNAi-resistant WT, S401A, or the S401D mutant form of APPL1. Transfected cells were serum starved and treated with adiponectin (1 μg/ml, 10 min). WT and mutant APPL1 were immunoprecipitated with an anti-myc antibody. Coimmunoprecipitated IRβ, IRS1, or IRS2 and their protein levels and phosphorylated APPL1 in the cell lysates were determined by western blot analysis using specific antibodies to these proteins.

(C) The model of the sensitization role of APPL1 in insulin signaling.

See also Figure S5.