Postreceptoral Adipocyte Insulin Resistance Induced by Nelfinavir Is Caused by Insensitivity of PKB/Akt to Phosphatidylinositol-3,4,5-Trisphosphate
Endocrinology Kachko et al. 150: 2618 Supplemental Data
Supplemental Data
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- Supplementary Figure 1 - Nelfinavir-induced increase in TRB3 does not mediate decreased insulin-stimulated PKB/Akt phosphorylation
A. 3T3-L1 adipocytes were treated with nelfinavir for 18 h, washed and stimulated with insulin for 10 min. Lysates were prepared and blotted with an anti-TRB3 antiserum (Calbiochem), anti-pSer473-PKB/Akt or anti-β actin (as loading control). The results indicate that nelfianvir-induced insulin resistance is associated with increased TRB3 expression.
B. and C. 24 h prior to nelfinavir treatment as above, 3T3-L1 adipocytes were electroporated in the presence of 2 nM siRNA directed to silence TRB3 (Ambion), or with a control siRNA sequence. B. shows representative blots, and C. quantification of insulin-stimulated phosphorylation of PKB/Akt. The results demonstrate effective prevention of nelfinavir-induced increase in TRB3 expression, yet, unaltered attenuation of insulin-stimulated Akt phosphorylation.