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. 2015 Jun 11;10(6):e0129644. doi: 10.1371/journal.pone.0129644

Fig 2. ZAG treatment impairs insulin action on glucose uptake and insulin signaling.

Fig 2

Differentiated human subcutaneous SGBS adipocytes (A) and visceral LiSa-2 adipocytes (B) were cultured for 24 hours with or without 25 μg/ml ZAG before stimulation with 100 nM insulin (Ins) for 30 minutes. Glucose uptake was measured during the final 10 min by incorporation of labelled 2-deoxyglucose into the cells. Left panels represent mean±SEM of 3–4 independent experiments performed in triplicate and are expressed as pmol glc/mg prot/10 min. Right panels represent percentage of stimulation produced by insulin over control cells (no insulin, without or with ZAG respectively). *, P < 0.01. (C) Lysates from differentiated SGBS cells and (D) LiSa-2 cells cultured with or without 25 μg/ml ZAG for 24 hours before stimulation with 100 nM insulin (Ins) for 15 minutes, were analyzed by western blotting using antibodies against phosphorylated and total IRβ (Tyr1150/1151), IRS1 (Tyr612), Akt (Ser473) and AS160 (Thr642). A representative experiment is shown together with densitometric analysis of phosphorylated vs total proteins (3 independent experiments). *, P < 0.01.