Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: The gene-specific primer pairs list in the study.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques:

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: Analysis of the mRNA expression of cytokines and transcription factors for Th cells in the colons of TRAF5 KO and WT mice. (a) TNF- α , (b) IFN- γ , (c) IL-4, (d) IL-17a, (e) IL-22, (f) T-bet, (g) GATA-3, (h) ROR- α , and (i) ROR- γ t mRNA expression levels were determined by real-time PCR. The data are representative of 3 independent experiments (mean ± SD). N = 8 per group. ∗ p < 0.05 versus the DSS-treated WT group; ∗∗ p < 0.01 versus the DSS-treated WT group. NS: not significant versus the DSS-treated WT group.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: Deletion of TRAF5 increases the protein expressions of the cytokines IFN- γ , IL-4, and IL-17a in the colons of mice induced by DSS. (a) The production of IFN- γ , IL-4, and IL-17a in the colons of DSS-induced mice or control mice was examined by ELISA ( N = 6 per group). (b) Representative immunofluorescence staining for IFN- γ , IL-4, and IL-17a in the colons of TRAF5 KO and WT mice (magnification 400x). (c) Quantification of positive cells ( N = 4 per group). The data are representative of 3 independent experiments (mean ± SD). ∗ p < 0.05 versus the DSS-treated WT group; ∗∗ p < 0.01 versus the DSS-treated WT group.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: Increased proportions of Th2 and IFN- γ /IL-17a-coproducing CD4 + T cells in the colons of TRAF5-deficient animals induced by DSS. LPMCs obtained from the colons of DSS-fed KO and WT mice were stimulated with PMA/ionomycin and subjected to surface staining for CD4. After fixation and permeabilization, intracellular staining for IL-4, IFN- γ , and IL-17a was performed as described in . CD4 + T cells were then gated and analyzed. (a) Representative flow plots. The percentages of (b) Th2 cells (CD4 + IL-4 + ), (c) IFN- γ /IL-17a-coproducing CD4 + T cells (CD4 + IFN- γ + IL-17a + ), (d) classical Th1 cells (CD4 + IFN- γ + IL-17a − ), and (e) classical Th17 cells (CD4 + IL-17a + IFN- γ − ) are presented as the mean ± SD. The data are representative of 3 independent experiments. N = 6 per group. ∗∗ p < 0.01 versus the DSS-treated WT group. NS: not significant versus the DSS-treated WT group.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques: Staining