Journal: eLife

Article Title: Thymic dendritic cell-derived IL-27p28 promotes the establishment of functional bias against IFN-γ production in newly generated CD4 + T cells through STAT1-related epigenetic mechanisms

doi: 10.7554/eLife.96868

Figure Lengend Snippet: ( A ) Intracellular staining of freshly isolated thymocytes from Cd11c-p28 f/f and WT mice using antibodies against phosphorylated STAT1 (Y701), STAT3 (Y705), and STAT4 (Y693). Representative histograms for CD4SP thymocytes (left) and mean fluorescence intensity (MFI) from three independent experiments (right, mean ± SD). ( B ) Western blot analysis of total and phosphorylated STAT1 (Y701), STAT3 (Y705), and STAT4 (Y693) in purified CD4SP thymocytes, CD4 + RTEs, and naive CD4 + T cells from Cd11c-p28 f/f and WT mice. Representative blots (left) and relative protein levels quantified by densitometry and normalization to β-actin (right, mean ± SD, n=3). ( C ) Increased STAT1 binding to promoter and regulatory regions of Tbx21 and Ifng loci in Cd11c-p28 f/f mice. ( D ) Correlation between STAT1 binding and H3K4me3 levels at Tbx21 and Ifng loci. Significance: * p <0.05; ** p <0.01. Figure 5—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 2. Original files for western blot analysis displayed in .

Article Snippet: Anti-phospho-STAT1(Tyr701), anti-phospho-STAT-1(S727), anti-STAT1, anti-phospho-STAT3 (Tyr705), anti-STAT3, anti-phospho-STAT4 (Tyr693), anti-STAT4, anti-SOCS3, anti-Actin and HRP-labeled goat-anti-rabbit or anti-mouse IgGs for western blot assay were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Staining, Isolation, Fluorescence, Western Blot, Purification, Binding Assay

Journal: PLOS One

Article Title: The ubiquitin-like modifier FAT10 does not affect IL-12 expression and signaling

doi: 10.1371/journal.pone.0323005

Figure Lengend Snippet: (A-D) CD4 + T cells were magnetically purified from spleens of C57BL/6 wild type mice or FAT10 -/- mice. (A, B) CD4 + T cells were treated with TNF (400 U/ml)/IFN-γ (200 U/ml) (indicated +) for 1 day or were left untreated (indicated -). pSTAT1, STAT1, and GAPDH were analyzed by western blot. (B) Quantification of pSTAT1 normalized to GAPDH. Data are depicted as mean ± SDs derived from 7 different experiments (n = 7). (C, D) CD4 + T cells were treated with TNF (400U/ml)/IFN-γ (200 U/ml) for 1 day or left untreated, followed by a treatment with IL-12 (10 ng/ml) for 2 h. Treatment regime is indicated. pSTAT4 and GAPDH were analyzed by western blot. (D) Quantification of pSTAT4 normalized to GAPDH. Data are depicted as mean ± SDs derived from 5 different experiments (n = 5).

Article Snippet: Protein was analyzed by the following antibodies: anti-pSTAT1 antibody (Cell Signaling, 7649S), anti-STAT1 antibody (Cell Signaling, 9172S), and anti-pSTAT4 antibody (Cell Signaling, 4134S).

Techniques: Purification, Western Blot, Derivative Assay