Ethylenediaminetetraacetic
acid (EDTA), p-nitrophenyl phosphate (pNPP), rosiglitazone, and DMSO were purchased from Sigma-Aldrich Co.
(St. Louis, MO). PTP1B (human recombinant) was purchased from Biomol
International LP (Plymouth Meeting, PA). Fetal bovine serum (FBS),
minimum essential medium (MEM), sodium pyruvate, penicillin–streptomycin,
and nonessential amino acids were purchased from Gibco-BRL Life Technologies
(Grand Island, NY). The fluorescentd -glucose analogue and
glucose tracer 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d -glucose (2-NBDG) was purchased from Life Technologies (Carlsbad,
CA). Human insulin was purchased from Eli Lilly (Fegersheim, France).
Phospho-Akt (Serine 473) (D9E) rabbit monoclonal antibody, p-IRS-1
(Tyr 895), and PEPCK antibody were obtained from Cell Signaling Technology
(Danvers, MA). PTP1B (D-4), IRS-1 (C-20) antibody, Akt1 (C-20), PI3-kinase
p110 (D-4), p-PI3-kinase p85α (Tyr 508), G6Pase-α (H-4),
β-Actin (C4), and all secondary antibodies were obtained from
Santa Cruz Biotechnology (Dallas, TX). The human hepatocarcinoma (HepG2)
cell line was purchased from the American Type Culture Collection
(HB-8065; Manassas, VA). All of the other chemicals and solvents used
were of reagent grade and were acquired from commercial sources.
acid (EDTA), p-nitrophenyl phosphate (pNPP), rosiglitazone, and DMSO were purchased from Sigma-Aldrich Co.
(St. Louis, MO). PTP1B (human recombinant) was purchased from Biomol
International LP (Plymouth Meeting, PA). Fetal bovine serum (FBS),
minimum essential medium (MEM), sodium pyruvate, penicillin–streptomycin,
and nonessential amino acids were purchased from Gibco-BRL Life Technologies
(Grand Island, NY). The fluorescent
glucose tracer 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-
CA). Human insulin was purchased from Eli Lilly (Fegersheim, France).
Phospho-Akt (Serine 473) (D9E) rabbit monoclonal antibody, p-IRS-1
(Tyr 895), and PEPCK antibody were obtained from Cell Signaling Technology
(Danvers, MA). PTP1B (D-4), IRS-1 (C-20) antibody, Akt1 (C-20), PI3-kinase
p110 (D-4), p-PI3-kinase p85α (Tyr 508), G6Pase-α (H-4),
β-Actin (C4), and all secondary antibodies were obtained from
Santa Cruz Biotechnology (Dallas, TX). The human hepatocarcinoma (HepG2)
cell line was purchased from the American Type Culture Collection
(HB-8065; Manassas, VA). All of the other chemicals and solvents used
were of reagent grade and were acquired from commercial sources.