Journal: Oxidative Medicine and Cellular Longevity

Article Title: IL-22 Protects against Biliary Ischemia-Reperfusion Injury after Liver Transplantation via Activating STAT3 and Reducing Apoptosis and Oxidative Stress Levels In Vitro and In Vivo

doi: 10.1155/2022/9635075

Figure Lengend Snippet: IL-22 plays a protective role by activating STAT3 in vitro. (a) Expression of STAT3 and p-STAT3 proteins in HIBEpiCs treated with IL-22 at different times. (b) Expression of STAT3 and p-STAT3 proteins after using 10 μ M and 20 μ M concentrations of STAT3 inhibitor stattic. (c) The cell cycle was detected after stattic (10 μ M) treatment, and there was no significant difference in the proportion of cells in the 4N phase compared with the CoCl 2 group. Data are shown as mean ± SD. ∗ P < 0.05. NS: no statistical difference.

Article Snippet: Subsequently, we treated the HIBEpiCs with the STAT3 inhibitor, stattic (MedChemExpress, USA), at concentrations of 10 μ M and 20 μ M for 48 h and detected the inhibition efficiency by western blot assay.

Techniques: In Vitro, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: IL-22 Protects against Biliary Ischemia-Reperfusion Injury after Liver Transplantation via Activating STAT3 and Reducing Apoptosis and Oxidative Stress Levels In Vitro and In Vivo

doi: 10.1155/2022/9635075

Figure Lengend Snippet: In vivo validation that IL-22 reduces IRI-induced apoptosis by activating STAT3. (a) The expression of STAT3 and p-STAT3 after intraperitoneal injection of RcIL-22 in rats for 1, 2, and 6 hours. (b) RcIL-22 can reduce the expression of cleaved-caspase3 and BAX protein and increase the expression of BCL2 and BCLXL protein in rats compared with the IRI group. (c) Evaluation of TUNEL expression in rat bile duct tissue sections by immunofluorescence staining. Data are shown as mean ± SD. ∗ P < 0.05.

Article Snippet: Subsequently, we treated the HIBEpiCs with the STAT3 inhibitor, stattic (MedChemExpress, USA), at concentrations of 10 μ M and 20 μ M for 48 h and detected the inhibition efficiency by western blot assay.

Techniques: In Vivo, Biomarker Discovery, Expressing, Injection, TUNEL Assay, Immunofluorescence, Staining

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: Frontiers in Immunology

Article Title: A Functionally Different Immune Phenotype in Cattle Is Associated With Higher Mastitis Incidence

doi: 10.3389/fimmu.2018.02884

Figure Lengend Snippet: Lymphocyte proteomes reveal regulation of differential expression of proteins between normal and deviant immune responders to PregSure BVD vaccination. Differential proteome analyses of PBL from control (Ctr, n = 2, shown as arithmetic means) and BNP ( n = 2, shown as arithmetic means) cows already revealed differences in protein expression patterns in constitutive expression (cE) of proteins, that shifted even stronger after (A) PWM and (B) ConA stimulation (48 h). Highly abundant proteins are presented in green and low abundant proteins in red. (C/D) In-depth analysis of part of the proteome relating to regulation of immune cells revealed some differentially (factor ≥ 2.0) expressed STAT proteins in bovine PBL after polyclonal activation with PWM (C) or ConA (D) . STAT3 and STAT5A were selectively upregulated in BNP lymphocytes (C) compared to controls due to PWM. After ConA stimulation, controls significantly upregulated STAT1 and STAT6 (D) , whereas in PBL of BNP cases STAT3 was enhanced during immune response (D) .

Article Snippet: For the inhibition assays, cells were incubated with STAT3 inhibitor (WP 1,066, Santa Cruz, Heidelberg, Germany, 50 ng/ml) 12 h before stimulation.

Techniques: Quantitative Proteomics, Control, Expressing, Activation Assay

Journal: Frontiers in Immunology

Article Title: A Functionally Different Immune Phenotype in Cattle Is Associated With Higher Mastitis Incidence

doi: 10.3389/fimmu.2018.02884

Figure Lengend Snippet: In PBL of control and BNP cows, different transcription factors were activated after ConA stimulation in vitro . (A) Representative western blot of constitutive (cE) pSTAT1 Tyr701 expression and after stimulation with ConA (ConA) in control PBL (Ctr, n = 1) and BNP PBL (BNP, n = 1). (B) Lymphocytes of vaccinated control cows (white-black striped bars, n = 5) phosphorylated STAT1 Tyr701 significantly stronger than BNP lymphocytes (black-white striped bars, n = 5) after ConA stimulation (** p < 0.01). (C) Representative western blot of constitutive (cE) pSTAT3 Tyr705 expression and after stimulation with ConA (ConA) in control PBL (Ctr, n = 1) and BNP PBL (BNP, n = 1). (D) BNP lymphocytes (black-white striped bars, n = 5) phosphorylated STAT3 Tyr705 comparatively stronger than vaccinated control lymphocytes (white-black striped bars, n = 5) in response to ConA stimulation (* p < 0.05). (A/C) Phosphorylated STAT1 Tyr701 and pSTAT3 Tyr705 signals were normalized to beta-actin and quantified using Image-J software. Protein expression is shown in mean ± SD. For statistical analyses Student‘s t test was performed.

Article Snippet: For the inhibition assays, cells were incubated with STAT3 inhibitor (WP 1,066, Santa Cruz, Heidelberg, Germany, 50 ng/ml) 12 h before stimulation.

Techniques: Control, In Vitro, Western Blot, Expressing, Software

Journal: Frontiers in Immunology

Article Title: A Functionally Different Immune Phenotype in Cattle Is Associated With Higher Mastitis Incidence

doi: 10.3389/fimmu.2018.02884

Figure Lengend Snippet: STAT3 inhibitor WP1066 selectively inhibits hyperproliferation of lymphocytes from BNP cows. STAT3 inhibitor did not inhibit proliferation of control PBL (white bar, n = 4), but significantly inhibited ConA stimulated BNP cells (black bars, n = 4, *** p < 0.001). Percentage of inhibition rate shown in mean ± SD. Student‘s t- test was used for statistical analyses.

Article Snippet: For the inhibition assays, cells were incubated with STAT3 inhibitor (WP 1,066, Santa Cruz, Heidelberg, Germany, 50 ng/ml) 12 h before stimulation.

Techniques: Control, Inhibition

Journal: Integrative Cancer Therapies

Article Title: Antitumor Efficacy of Huqizhengxiao (HQZX) Decoction Based on Inhibition of Telomerase Activity in Nude Mice of Hepatocarcinoma Xenograft

doi: 10.1177/1534735418785999

Figure Lengend Snippet: Influence of HQZX decoction on signaling molecules of HepG2 xenograft. Fold changes of p-ERK/ERK, p-STAT3/STAT3 and p-mTOR/mTOR of treatment groups versus control group. * P < .05 compared with control group.

Article Snippet: Total/phosph ERK MAG Kit, total/phosph STAT3 MAG Kit, total/phosph mTOR MAG Kit and tubulin MAG kit were from Merck Millipore.

Techniques: Control