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TNF-α and insulin act synergistically to promote phosphorylation of Ser307 in IRS-1. The 3T3-L1 preadipocytes were treated for 30 minutes with 20 ng/ml TNF-α and 100 nM insulin either separately or simultaneously. IRS-1 in cell lysates was immunoprecipitated with αIRS-1 and immunoblotted with αpS307.
