Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Disease Risk-Associated Genetic Variants in STAT1 and STAT4 Function in a Complementary Manner to Increase Pattern-Recognition Receptor–Induced Outcomes in Human Macrophages

doi: 10.4049/jimmunol.1901112

Figure Lengend Snippet: STAT1 and STAT4 are required for optimal secretion of cytokines in MDMs upon stimulation through a broad range of PRRs. (A, C, and D) Human MDMs were transfected with scrambled siRNA or with STAT1 or STAT4 siRNA, alone or in combination, and then treated for 24 h with the following: (A) 0.1 μg/ml lipid A (TLR4) (n = 6, similar results seen in an independent n = 8), (C) 10 μg/ml Pam3Cys (TLR2), 100 μg/ml poly(I:C) (TLR3), or 10 μg/ml CpG DNA (TLR9) (n = 6, similar results seen in an independent n = 8), or (D) S. Typhimurium (n = 6). (B) MDMs were pretreated for 1 h with a STAT1 inhibitor (fludarabine) or a STAT4 inhibitor (lisofylline), alone or in combination, or with vehicle (DMSO) and then with 0.1 μg/ml lipid A for 24 h (n = 4). (A–D) Mean cytokine secretion + SEM. Significance is shown compared with stimulated, scrambled siRNA–transfected cells or stimulated, vehicle control-treated cells. t test with Bonferroni–Holm correction. **p < 0.01, ***p < 0.001, †p < 1 × 10−4, ††p < 1 × 10−5. scr, scrambled; Tx, treatment.

Article Snippet: In some cases, cells were pretreated with 5 μg/ml fludarabine (STAT1 inhibitor), 150 μM lisofylline (STAT4 inhibitor) (Cayman Chemical Company), Upadacitinib (JAK1 inhibitor), or BMS-986165 (TYK2 inhibitor) (MedChemExpress).

Techniques: Transfection, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Disease Risk-Associated Genetic Variants in STAT1 and STAT4 Function in a Complementary Manner to Increase Pattern-Recognition Receptor–Induced Outcomes in Human Macrophages

doi: 10.4049/jimmunol.1901112

Figure Lengend Snippet: STAT1 and STAT4 promote S. Typhimurium–induced cytokine secretion in human intestinal myeloid cells. Human intestinal myeloid cells (n = 6 donors) or peripheral MDMs (n = 6 donors) were preincubated for 1 h with either fludarabine (STAT1 inhibitor) or lisofylline (STAT4 inhibitor), alone or in combination, and then cocultured with S. Typhimurium (S. Typhim) at MOI 10:1 for 24 h. Cytokine secretion + SEM. Significance with t test with Bonferroni–Holm correction. ***p < 0.001, †p < 1 × 10−4, ††p < 1 × 10−5. Inh, inhibitor.

Article Snippet: In some cases, cells were pretreated with 5 μg/ml fludarabine (STAT1 inhibitor), 150 μM lisofylline (STAT4 inhibitor) (Cayman Chemical Company), Upadacitinib (JAK1 inhibitor), or BMS-986165 (TYK2 inhibitor) (MedChemExpress).

Techniques:

Journal: Scientific Reports

Article Title: IL-17 Promotes Angiogenic Factors IL-6, IL-8, and Vegf Production via Stat1 in Lung Adenocarcinoma

doi: 10.1038/srep36551

Figure Lengend Snippet: (A , B) A549/H292 cells cells (1 × 10 6 cells/well) were treated with or without 100 ng/ml IL-17 for 6 h, and then STAT1 protein levels were determined by WB. Target gene expression was normalized to the Gapdh housekeeping gene, and the data from three independent experiments are presented. (C) A549/H292 cells cells (5 × 10 5 cells/well) were treated with or without 100 ng/ml IL-17 for 6 h, and then STAT1 mRNA levels were determined by qRT-PCR. mRNA expression levels were calculated using the 2 −ΔΔCt method, and target gene expression was normalized to the GAPDH housekeeping gene. The results shown are representative of four independent experiments and are presented the mean ± SEM. *p < 0.05 was regarded as significant. *Means p < 0.05, **means p < 0.01, ***means p < 0.001.

Article Snippet: The STAT1 inhibitor (fludarabine) was obtained from HISUN Pharmaceutical Co. Ltd. (Zhejiang, China).

Techniques: Targeted Gene Expression, Quantitative RT-PCR, Expressing

Journal: Scientific Reports

Article Title: IL-17 Promotes Angiogenic Factors IL-6, IL-8, and Vegf Production via Stat1 in Lung Adenocarcinoma

doi: 10.1038/srep36551

Figure Lengend Snippet: A549/H292 cells were incubated with IL-17 or IL-17 plus a STAT1 inhibitor for 6 or 48 h (100 ng/ml IL-17; and 30 μM inhibitor). The IL-6, IL-8, and VEGF mRNA and protein levels were determined by qRT-PCR and ELISA, respectively. (A) IL-6, IL-8, and VEGF mRNA levels in A549/H292 cells, mRNA expression levels were calculated using the 2 −ΔΔCt method, and target gene expression was normalized to the GAPDH housekeeping gene. The data are presented as the mean ± SEM of three independent experiments. (B) IL-6, IL-8, and VEGF protein levels in A549/H292 cells. The data are presented as the mean ± SEM of three independent experiments. Comparisons were performed using the t-test. *p < 0.05; **p < 0.01; and ***p < 0.001.

Article Snippet: The STAT1 inhibitor (fludarabine) was obtained from HISUN Pharmaceutical Co. Ltd. (Zhejiang, China).

Techniques: Incubation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Targeted Gene Expression

Journal: Scientific Reports

Article Title: IL-17 Promotes Angiogenic Factors IL-6, IL-8, and Vegf Production via Stat1 in Lung Adenocarcinoma

doi: 10.1038/srep36551

Figure Lengend Snippet: A549/H292 cells (2 × 10 5 cells/well) were transfected with STAT1 siRNA for 48 h. (A) STAT1 protein expression was determined by WB. Then, A549/H292 cells (5 × 10 5 cells/well) were transfected with siRNA for 48 h and subsequently treated with human IL-17 (100 ng/ml) for an additional 6 or 48 h. The IL-6, IL-8, and VEGF mRNA and protein expression levels were determined by qRT-PCR and WB, respectively. (B) IL-6, IL-8, VEGF mRNA levels in A549/H292 cells. mRNA expression levels were calculated using the 2 −ΔΔCt method, and target gene expression was normalized to the GAPDH housekeeping gene. The data are presented as the mean ± SEM of three independent experiments. (C) IL-6, IL-8, and VEGF protein levels in A549/H292 cells and the results are presented as the mean ± SEM of three independent experiments. Comparisons were performed using the t-test. *p < 0.05; **p < 0.01; and ***p < 0.001.

Article Snippet: The STAT1 inhibitor (fludarabine) was obtained from HISUN Pharmaceutical Co. Ltd. (Zhejiang, China).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Targeted Gene Expression

Journal: Scientific Reports

Article Title: IL-17 Promotes Angiogenic Factors IL-6, IL-8, and Vegf Production via Stat1 in Lung Adenocarcinoma

doi: 10.1038/srep36551

Figure Lengend Snippet: (A , B) The relative protein expression of IL-6, IL-8, VEGF and STAT1 in tumour tissues of A549-IL-17 vs. A549-Neo cell-bearing nude mice was determined by WB. (C) The relative mRNA expression of IL-6, IL-8, VEGF and STAT1 in tumour tissues of A549-IL-17 vs. A549-Neo cell-bearing nude mice was determined by qRT-PCR (n = 5). mRNA expression levels were calculated using the 2 −ΔΔCt method, and target gene expression was normalized to the GAPDH housekeeping gene. The data are presented as the mean ± SEM for five mice per group, and the results are representative of two independent experiments. *p < 0.05; **p < 0.01; and ***p < 0.001.

Article Snippet: The STAT1 inhibitor (fludarabine) was obtained from HISUN Pharmaceutical Co. Ltd. (Zhejiang, China).

Techniques: Expressing, Quantitative RT-PCR, Targeted Gene Expression