Journal: Journal for Immunotherapy of Cancer

Article Title: Type I interferon suppresses tumor growth through activating the STAT3-granzyme B pathway in tumor-infiltrating cytotoxic T lymphocytes

doi: 10.1186/s40425-019-0635-8

Figure Lengend Snippet: IFNα and IFNβ activate STAT3 to up-regulate Gzmb expression in CTLs. a . the tumor-specific resting 2/20 CTLs were cultured in the presence of IFNα and IFNβ, respectively, and analyzed by Western blotting analysis for the indicated STATs. b . The protein band intensities of pSTAT1 and pSTAT3 as shown in A were quantified using NIH image J and normalized as the ratio over the intensities of STAT1 and STAT3, respectively. Column: Mean; Bar: SD. c . Resting 2/20 CTLs were treated with recombinant IFNα and IFNβ, respectively, in the absence (control) or presence of pSTAT1 (+Fludarabine, 10 μM, top panel) and pSTAT3 (+STATTIC, 5 μM, bottom panel) inhibitors, respectively, for 24 h. Cells were analyzed by qPCR for Gzmb expression level

Article Snippet: Fluorescent dye-conjugated antibodies that are specific for CD45, CD4, CD8, and Zombie violet were obtained from Biolegend (San Diego, CA). pSTAT1 inhibitor Fludarabine [ ] and pSTAT3 inhibitor Stattic [ ] were obtained from Santa Cruz.

Techniques: Expressing, Cell Culture, Western Blot, Recombinant, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: Type I interferon suppresses tumor growth through activating the STAT3-granzyme B pathway in tumor-infiltrating cytotoxic T lymphocytes

doi: 10.1186/s40425-019-0635-8

Figure Lengend Snippet: IFNα and IFNβ-activated STAT3 binds to the Gzmb promoter in CTLs. a . Structures of the Gzmb promoter. The six putative ISRE sequences (right panel) and locations (left panel) are shown. b . Resting 2/20 CTLs were treated with recombinant IFNα and IFNβ protein, respectively, for 1 h. Nuclear extracts were prepared from these cells and analyzed for STAT3 activation using EMSA with the WT pSTAT3 consensus probe (Santa Cruz Cat# sc-2571) and mutant probe (Santa Cruz Cat# sc-2572). Black arrow points to the DNA-pSTAT3 complex. c . Nuclear extracts were prepared as in B and analyzed for STAT3 activation using EMSA with the Gzmb promoter DNA probe GP4 as indicated in A. To determine pSTAT3-DNA binding specificity, the WT pSTAT3 consensus probe as shown in B was used for cold probe competition at the indicated ratios relative to the GP4 probe as a specificity control. Black arrow points to the DNA-pSTA T3 complex

Article Snippet: Fluorescent dye-conjugated antibodies that are specific for CD45, CD4, CD8, and Zombie violet were obtained from Biolegend (San Diego, CA). pSTAT1 inhibitor Fludarabine [ ] and pSTAT3 inhibitor Stattic [ ] were obtained from Santa Cruz.

Techniques: Recombinant, Activation Assay, Mutagenesis, Binding Assay, Control

Journal: Journal of Inflammation Research

Article Title: Anti-Inflammatory Regulatory Role of Signal Transducer and Activator of Transcription 3 Phosphorylation in Regulating Hypersensitivity Responses to Echinococcus granulosus Hydatid Cyst Fluid

doi: 10.2147/JIR.S509286

Figure Lengend Snippet: The inhibitory effect of different concentrations of inhibitors on STAT3 phosphorylation.

Article Snippet: The STAT3 inhibitors Stattic and JSI-124 were procured from Shanghai Haoyuan Chemexpress Co., Ltd. Antibodies for STAT3, phosphorylated p-STAT3 (S727), p-STAT3 (Y705), and ACTIN, as well as secondary antibodies, were purchased from Wuhan ABclonal Biotechnology Co., Ltd. High-resolution rapid electrophoresis liquid and ice-free rapid transfer liquid were obtained from Wuhan Servicebio Technology Co., Ltd. Immunoglobulin E (IgE) was purchased from Sigma-Aldrich, USA.

Techniques: Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Resveratrol Is a Natural Inhibitor of Human Intestinal Mast Cell Activation and Phosphorylation of Mitochondrial ERK1/2 and STAT3

doi: 10.3390/ijms22147640

Figure Lengend Snippet: Phosphorylation of signal transducer and activator of transcription 3 (STAT3) and extracellular signal-regulated kinase 1/2 (ERK1/2) in hiMC following treatment with stattic, nobiletin, and resveratrol. Western blot analyses of ( A ) phosphorylated STAT3 ( n = 5) and ( B ) phosphorylated ERK1/2 ( n = 4) in hiMC. HiMCs were treated with 60 μM of stattic (S), 45 μM of nobiletin (N), 50 μM of resveratrol (R), or corresponding concentrations of the vehicle DMSO (control) before stimulation by FcεRI crosslinking using 100 ng/mL of mAb 22E7 for 5 min. Representative pictures and densitometric analyses in relation to stimulated control are shown. Values are mean ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: To analyze STAT3 activation, hiMCs were pre-incubated for 20 min with 60 μM STAT3 inhibitor stattic (Merck) prior to stimulation by FcεRI crosslinking.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Resveratrol Is a Natural Inhibitor of Human Intestinal Mast Cell Activation and Phosphorylation of Mitochondrial ERK1/2 and STAT3

doi: 10.3390/ijms22147640

Figure Lengend Snippet: Phosphorylation of STAT3 and ERK1/2 in nuclear and mitochondrial fractions of hiMC following treatment with resveratrol. Western blot analyses of two independent experiments of ( A ) phosphorylated ERK1/2 and ( B ) phosphorylated STAT3 after subcellular fractionation in pure mitochondria (Mito), cytosol fraction (Cyto), and crude nuclear fraction (Nuc), respectively. HiMCs were treated with 50 μM of resveratrol (R), or corresponding concentrations of the vehicle DMSO (control), and stimulated by FcεRI crosslinking with 100 ng/mL of mAb 22E7 for 5 min. Exemplary pictures and densitometric analyses in relation to stimulated control for Western blots of phosphorylated ERK1/2 and total ERK1/2, and phosphorylated STAT3 and total STAT3, are shown. Values are mean ± SEM ( n = 2).

Article Snippet: To analyze STAT3 activation, hiMCs were pre-incubated for 20 min with 60 μM STAT3 inhibitor stattic (Merck) prior to stimulation by FcεRI crosslinking.

Techniques: Western Blot, Fractionation

Journal: Frontiers in Pharmacology

Article Title: Tirbanibulin Attenuates Pulmonary Fibrosis by Modulating Src/STAT3 Signaling

doi: 10.3389/fphar.2021.693906

Figure Lengend Snippet: The impact of KX-01 on pulmonary protein expression. (A, B) : The expression of HIF-1α, p-Src; (C, D) : p-STAT3, STAT3 and p-STAT3/STAT3 was analyzed. BLM: bleomycin. Data are means ± SD ( n = 3). # p < 0.01 vs. sham; * p < 0.01 vs. BLM. Data were compared via ANOVAs with Dunnett’s test.

Article Snippet: The STAT3 inhibitor Stattic (purity>98%, C 8 H 5 NO 4 S, CAS No: 19983-44-9) was from the Jiangsu Aikang Biomedicine Company (China).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Tirbanibulin Attenuates Pulmonary Fibrosis by Modulating Src/STAT3 Signaling

doi: 10.3389/fphar.2021.693906

Figure Lengend Snippet: The impact of KX-01 on pulmonary α-SMA and p-STAT3 immunohistochemical staining. (A) and (4C) : Immunohistochemical staining was used to assess pulmonary α-SMA; (B–D) : Immunohistochemical staining was used to assess pulmonary p-STAT3 expression following BLM treatment with or without KX-01 administration. BLM: bleomycin. Data are means ± SD. # p < 0.01 vs. sham; * p < 0.01 vs. BLM. Data were compared via ANOVAs with Dunnett’s test.

Article Snippet: The STAT3 inhibitor Stattic (purity>98%, C 8 H 5 NO 4 S, CAS No: 19983-44-9) was from the Jiangsu Aikang Biomedicine Company (China).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Frontiers in Pharmacology

Article Title: Tirbanibulin Attenuates Pulmonary Fibrosis by Modulating Src/STAT3 Signaling

doi: 10.3389/fphar.2021.693906

Figure Lengend Snippet: The impact of KX-01 on L929 cell proliferation and protein expression. (A) An EdU incorporation assay was used to assess L929 cell proliferation following CoCl 2 treatment with or without KX-01 for 72 h. (B–C) : The expression of α-SMA, collagen I, and collagen III were assessed; (D, E) : The expression of HIF-1α, p-Src (D–E); (F, G) : The expression of p-STAT3 and STAT3, and the ratio of p-STAT3/STAT3 was assessed. Data are means ± SD ( n = 3). # p < 0.01 vs. control; * p < 0.01 vs. CoCl 2 . Data were compared via ANOVAs with Dunnett's tst.

Article Snippet: The STAT3 inhibitor Stattic (purity>98%, C 8 H 5 NO 4 S, CAS No: 19983-44-9) was from the Jiangsu Aikang Biomedicine Company (China).

Techniques: Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Tirbanibulin Attenuates Pulmonary Fibrosis by Modulating Src/STAT3 Signaling

doi: 10.3389/fphar.2021.693906

Figure Lengend Snippet: The impact of KX-01 on L929 cell proliferation and protein expression in the presence of Roxadustat. (A) An EdU incorporation assay was used to assess L929 cell proliferation following CoCl 2 treatment with or without KX-01 or ROT for 72 h. (B, C) : The expression of α-SMA, collagen I, and collagen III were assessed; (D, E) : The expression of HIF-1α, p-Src, p-STAT3 and STAT3, and the ratio of p-STAT3/STAT3 were assessed. ROT: Roxadustat. Data are means ± SD ( n = 3). # p < 0.05, ## p < 0.01 vs. control; * p < 0.05, ** p < 0.01 vs. CoCl 2 . Data were compared via ANOVAs with Dunnett's test.

Article Snippet: The STAT3 inhibitor Stattic (purity>98%, C 8 H 5 NO 4 S, CAS No: 19983-44-9) was from the Jiangsu Aikang Biomedicine Company (China).

Techniques: Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Tirbanibulin Attenuates Pulmonary Fibrosis by Modulating Src/STAT3 Signaling

doi: 10.3389/fphar.2021.693906

Figure Lengend Snippet: The impact of KX-01 on L929 cell proliferation and protein expression in the presence of Stattic. (A) An EdU incorporation assay was used to assess L929 cell proliferation following CoCl 2 treatment with or without KX-01 or Stattic for 72 h. (B, C) : The expression of α-SMA, collagen I, and collagen III were assessed; (D, E) : The expression of HIF-1α, p-Src, p-STAT3 and STAT3, and the ratio of p-STAT3/STAT3 were assessed. Data are means ± SD ( n = 3). # p < 0.01 vs. control; * p < 0.05, ** p < 0.01 vs. CoCl 2 . Data were compared via ANOVAs with Dunnett's test.

Article Snippet: The STAT3 inhibitor Stattic (purity>98%, C 8 H 5 NO 4 S, CAS No: 19983-44-9) was from the Jiangsu Aikang Biomedicine Company (China).

Techniques: Expressing, Control