expression vectors socs3  (STARR Life Sciences)

Journal: Journal of Biological Chemistry

Article Title: Regulation of Interleukin-6-induced Hepatic Insulin Resistance by Mammalian Target of Rapamycin through the STAT3-SOCS3 Pathway

doi: 10.1074/jbc.m708568200

Figure Lengend Snippet: FIGURE 7. A Jak inhibitor rescues insulin signaling from IL-6 inhibition. A, serum-starved HepG2 cells were stimulated by 20 ng/ml IL-6 with or with- out 100 nM rapamycin for 1 h. Relative levels of SOCS3 mRNA were deter- mined by quantitative RT-PCR. The average results of three independent experimentsareshownwitherrorbarsrepresentingS.D.*,p0.01compared with IL-6 stimulation in the absence of rapamycin (Student’s t test). B, serum- starved HepG2 cells were stimulated by 20 ng/ml IL-6 for 30 or 45 min with or without 10 M AG490. Relative levels of SOCS3 mRNA were determined by quantitative RT-PCR. The average results of three independent experiments are shown with error bars representing S.D. *, p 0.05 when comparing data with and without AG490 treatment at each IL-6 stimulation time point (Stu- dent’s t test). C, serum-starved cells were stimulated with 100 nM insulin for 10 min, with or without pretreatment by 20 ng/ml IL-6 for 2.5 h. When indicated, 100 nM rapamycin or 10 M AG490 was added 30 min prior to stimulation. Cell lysatesweresubjectedtoWesternanalysesforphospho-Akt(Ser473)andtotal Akt. Rap, rapamycin.

Article Snippet: Human SOCS3 cDNA (in pCMV6-XL6) was obtained from Origene and subcloned into pCDNA3-Myc via NotI.

Techniques: Inhibition, Quantitative RT-PCR