Fig. 7

Analysis of SOCS3 expression and STAT1/3 phosphorylation following treatment with IL-27, sIL-6R/IL-6 or both cytokines. a sIL-6R/IL-6 (hy-IL-6) or IL-27, either alone or in combination, mediates only partial changes of SOCS3 mRNA expression by QRT-PCR. Constitutive expression of mRNA is also evident. Data, normalized to GAPDH housekeeping gene, are expressed as 1/ΔCT. Error bars represent SD in one representative experiment out of two with consistent data. b Western blot analysis reveals constitutive SOCS3 expression (untreated control sample, CTR), which is marginally effected by sIL-6R/IL-6 (hy-IL-6) stimulation, at the indicated time intervals, in NCI-H69 cells. Tyrosine phosphorylated (P)-STAT1 and P-STAT3 proteins are analyzed as control of signal transduction. c Western blot analysis of NCI-H69 cells treated for 30 min with the indicated cytokines shows that IL-27 mediates stronger induction of P-STAT1, which is not inhibited by concomitant stimulation with sIL-6R/IL-6 (hy-IL-6). Tubulin served as loading control