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. 2015 Dec 11;37(2):347–358. doi: 10.3892/ijmm.2015.2433

Figure 3.

Figure 3

Nuclear translocation and nitric oxide synthase 2 (NOS2) promoter binding of Tyr701-phosphorylated signal transducer and activator of transcription 1 [p-STAT1(Y701)] by interferon-γ (IFN-γ) and/or interleukin-17 (IL-17) in RAW 264.7 cells. (A-a) Representative western blot analysis of nuclear translocation of p-STAT1(Y701) in RAW 264.7 cells treated with the indicated cytokines for 1 h. β-actin was used as a cytoplasmic loading control, and histone H3 was used as a nuclear loading control. (A-b) Average quantification of nuclear and cytoplasmic p-STAT1(Y701) obtained by densitometric analysis for western blot analysis. Data are expressed as the density ratio of target protein to its non-treated level in arbitrary units. (A-c) p-STAT1(Y701) ratio of nucleus to cytoplasm. (B) Chromatin immunoprecipitation (ChIP) analysis of p-STAT1(Y701) occupancy on the proximal interferon-gamma-activated site (GAS) of NOS2 promoter in RAW 264.7 cells treated with indicated cytokines for 1 h. Antibodies: R, anti-RNA polymerase II antibody; I, IgG; S, anti-p-STAT1(Y701) antibody. Data are presented as the means ± SD from three independent experiments. *p<0.05 and **p<0.01.