Journal: Respiratory Research

Article Title: IL-35 subunit EBI3 alleviates bleomycin-induced pulmonary fibrosis via suppressing DNA enrichment of STAT3

doi: 10.1186/s12931-021-01858-x

Figure Lengend Snippet: IL-35 inhibited ECM-associated genes via competitively identifying STAT3 binding sites. A Lung epithelial cells were stimulated with IL-35 (5 ng/ml) for indicated time points, and phosphorylation of STAT1 and STAT4 were measured by WB. B Lung epithelial cells were co-stimulated with IL-35 (5 ng/ml) and IL-6 (10 ng/ml), and phosphorylation of STAT3 were measured by WB. C CHIP qPCR assay for evaluating the enrichment of STAT1 and STAT4 on the promoters of Col1a2 and Col3a1. Input was used as the relative normalization (n = 3). Data are representative of three independent experiments. Data are presented as mean ± SEM. Significance was determined by two-tailed Student’s t test. *P < 0.05

Article Snippet: Antibodies used in this study were listed below: rabbit monoclonal antibody for phospho-Stat1 (Tyr701) (D4A7, 1:1000 dilution), Stat1 (D1K9Y, 1:1000 dilution), phospho-Stat3 (Tyr705) (D3A7, 1:1000 dilution), Stat3 (D3Z2G, 1:1000 dilution) and Stat4 (C46B10, 1:1000 dilution) were purchased from Cell Signaling Technology (Danvers, USA).

Techniques: Binding Assay, Phospho-proteomics, ChIP-qPCR, Two Tailed Test

Journal: iScience

Article Title: Extracellular nicotinamide phosphoribosyltransferase boosts IFNγ-induced macrophage polarization independently of TLR4

doi: 10.1016/j.isci.2022.104147

Figure Lengend Snippet:

Article Snippet: Antibodies used were as follows: mouse (Mo) anti-NAMPT from AdipoGen (OMNI379); rabbit (Rb) anti-NAMPT GTX128973 from GeneTex; Mo anti-βactin A1978 from Sigma, Rb anti-phospho-STAT3 (Tyr705) (D3A7) from Cell Signaling, mo anti-STAT3 (124H6) from Cell Signaling, Rb anti-phospho-STAT1 (Tyr701) (D4A7) from Cell Signaling and Rb anti-STAT1 (D1K9Y) from Cell Signaling.

Techniques: Staining, Virus, Recombinant, Bradford Protein Assay, Endotoxin Assay, Enzyme-linked Immunosorbent Assay, Software

Journal: Virology journal

Article Title: RUNX1 inhibits the antiviral immune response against influenza A virus through attenuating type I interferon signaling.

doi: 10.1186/s12985-022-01764-8

Figure Lengend Snippet: Fig. 5 RUNX1 attenuated IRF3 and STAT1 signaling. a The mRNA level of TRAF3, RIG-I, MAVS, TBK1, IRF3, and STAT1 in the shRUNX1 and shControl cells were assessed by qRT-PCR. b The mRNA of these genes in A549 cells that were transfected with empty plasmid pCMV-GFP or pCMV-RUNX1 was assessed by qRT-PCR. c, d The protein and the phosphorylation level of IRF3 and STAT1 of these cells were assessed by Western blot. e, f These cells were infected with PR8 (MOI = 5) and collected at 3, 6, and 9 h.p.i. The protein and the phosphorylation levels of IRF3 and STAT1 in these cells were assessed by Western blot. Data are mean ± SD of three independent experiments. Significance is by unpaired T-test; *p < 0.05; **p < 0.01

Article Snippet: Mouse anti-RUNX1(A-2) (sc-365644) antibodies were purchased from Santa Cruz, Dallas, TX, USA; rabbit antiIRF3 (11312-AP) and rabbit anti-STAT1 (10144–2-AP) were purchased from Proteintech, Rosemont, IL, USA; rabbit anti-P-IRF3 (Ser396) (4D4G) antibodies and rabbit anti-P-STAT1 (Tyr701) (D4A7) antibodies were purchased from Cell Signaling, Danvers, MA, USA; mouse anti-GAPDH antibody (AF0006) was purchased from Beyotime, Shanghai, China; mouse mAbs to viral proteins NP and M1 of IAV were obtained from Dr. Jiyong Zhou [35].

Techniques: Quantitative RT-PCR, Transfection, Plasmid Preparation, Phospho-proteomics, Western Blot, Infection

Journal: Journal for Immunotherapy of Cancer

Article Title: Type I T cells sensitize treatment refractory tumors to chemotherapy through inhibition of oncogenic signaling pathways

doi: 10.1136/jitc-2021-002355

Figure Lengend Snippet: IFN-γ suppresses signaling through multiple oncogenic growth factor receptors in tumor cells. (A) Western blots of the indicated proteins from MMC cells unstimulated and after IFN-γ treatment with tubulin loading control. Densitometric quantification of the (B) indicated growth factor receptor or (C) signaling molecule after treatment with IFN-γ (blue bars) compared with the untreated control cells (gray bars); n=3 independent experiments, **P<0.01, ***P<0.001, ****P<0.0001. IFN-γ, interferon gamma; IR, insulin receptor; p-IR, phosphorylated IR; IGF-IR, insulin-like growth factor receptor-1; p-IGF-IR, phosphorylated IGF-IR; EGFR, epidermal growth factor receptor; p-EGFR, phosphorylated EGFR; PTEN, phosphatase and tensin homolog; AKT, serine/threonine-protein kinase; p-AKT, phosphorylated AKT; STAT-1, signal transducer and activator of transcription 1; p-STAT-1, phosphorylated STAT-1.

Article Snippet: Antibodies used were phospho-IGF-IR (Tyr 1161, #sc-101703), IGF-IRα (clone, N-20), phospho-epidermal growth factor receptor (EGFR) (Tyr 1173, #sc-12351), EGFR (clone 1005), phospho-insulin receptor (IR) β (clone 10C3), IR β (clone C-19) all at 1 μg/mL (Santa Cruz Biotechnology), α-tubulin (clone B-7), phospho-signal transducer and activator of transcription (STAT-1; Tyr 701, clone D4A7), STAT-1 (clone D1K9Y), non-phospho-phosphatase and tensin homolog (PTEN;Ser380/Thr382/Thr383, #9569), SOCS1 (clone A156), SOCS2 (#2779), phospho-serine/theronine-protein kinase (AKT) (Ser473, #9271) and AKT (#9272), all at 1 μg/mL (Cell Signaling Technology), SOCS3 (#626602) at 1 μg/mL (Biolegend) and horseradish peroxidase-conjugated goat anti-rabbit (20 ng/mL, Thermo Fisher Scientific).

Techniques: Western Blot, Control