Journal: Molecular Cancer

Article Title: ISL-1 is overexpressed in non-Hodgkin lymphoma and promotes lymphoma cell proliferation by forming a p-STAT3/p-c-Jun/ISL-1 complex

doi: 10.1186/1476-4598-13-181

Figure Lengend Snippet: JNK or JAK/STAT signaling inhibitors inhibit NHL cells proliferation through down-regulating ISL-1 expression. (A) The relative proliferation rate of lymphoma cell lines were measured using CCK-8 analysis after treated with JNK signaling pathway inhibitor (SP60012, 10 μM) or JAK/STAT signaling pathway inhibitor (STATTIC, 6 μM). The cell treated with DMSO were used as the control. (B to C) The effect of SP600125 (10 μM) and STATTIC (6 μM) on ISL-1 and c-Myc expression at both mRNA (B) and protein levels (C) were analyzed by real-time RT-PCR and Western blot. The cells treated with DMSO at different time point were used as the corresponding control. (D) The luciferase activity of c-Myc-luc (wide type or M1) was measured in Ly3 cells with or without ISL-1 transcfection after treated with 10 μM SP600125 or 6 μM STATTIC for 24 h. (E) The growth inhibition of Ly3 cells with or without ISL-1 transcfection was measured by CCK-8 analysis after treated with 10 μM SP600125 or 6 μM STATTIC for 24 h. The data represent three independent experiments. Each bar represents mean ± SD. p values were calculated using a Student t -test (* p < 0.05, ** p < 0.01, # p < 0.05, ## p < 0.01 vs. each control).

Article Snippet: STATTIC (sc-202818, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SP600125 (s5567) and Anisomycin (A9789) purchased from Sigma (Louis, MO, USA) were dissolved in 100% dimethyl sulfoxide to prepare a 40 mM stock, 20 mM stock and 15 mg/ml stock respectively, and stored at -20°C.

Techniques: Expressing, CCK-8 Assay, Control, Quantitative RT-PCR, Western Blot, Luciferase, Activity Assay, Inhibition

Journal: Molecular Cancer

Article Title: ISL-1 is overexpressed in non-Hodgkin lymphoma and promotes lymphoma cell proliferation by forming a p-STAT3/p-c-Jun/ISL-1 complex

doi: 10.1186/1476-4598-13-181

Figure Lengend Snippet: p-STAT3/p-c-Jun/ ISL-1 forms a transcriptional complex and binds directly to ISL-1 promoter. (A) Consensus binding sites for p-STAT3 and p-c-Jun on the ISL-1 promoter were analyzed by Matinspector software. (B) The luciferase activity of ISL-1-luc was analyzed by luciferase reporter assay in Ly3 cells after treated with IL-6 (4 ng/ml), STATTIC (6 μM), Anisomycin (15 ng/ml) or SP600125 (10 μM) for 24 h. (C) ChIP assay was performed with anti-p-STAT3 Ab (left panel) or anti-p-c-Jun Ab (right panel) for immunoprecipitation using chromatin harvested from Ly3 cells. The DNA extractions were amplified using the primers that cover the p-STAT3 (primers 2) or p-c-Jun (primers 4) binding sites, or control primers (primers 1, 3) on the ISL-1 promoter by real-time PCR with normal IgG as a control. (D) Co-IP assay was performed in Ly3 for the transcriptional complex recruited on the ISL-1 promoter. (E) ChIP-re-IP assay was performed first with anti-ISL-1 Ab or rabbit IgG Ab and then with anti-p-STAT3, anti-p-c-Jun or IgG Abs for immunoprecipitation using chromatin harvested from Ly3 cells. (F) The transcriptional activity of ISL-1 on ISL-1-luc was analyzed in Ly3 cells by luciferase reporter assay. The data represent three independent experiments. Each bar represents mean ± SD. p values were calculated using a Student t -test (* p < 0.05, ** p < 0.01 vs. the control).

Article Snippet: STATTIC (sc-202818, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SP600125 (s5567) and Anisomycin (A9789) purchased from Sigma (Louis, MO, USA) were dissolved in 100% dimethyl sulfoxide to prepare a 40 mM stock, 20 mM stock and 15 mg/ml stock respectively, and stored at -20°C.

Techniques: Binding Assay, Software, Luciferase, Activity Assay, Reporter Assay, Immunoprecipitation, Amplification, Control, Real-time Polymerase Chain Reaction, Co-Immunoprecipitation Assay

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: The effect of tumor microenvironment on activities of M1 macrophages and M1 macrophages with inhibited transcription factors, STAT3 and STAT6.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques:

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Changes in the ratio of mean% changes in all M1 cytokines to mean% changes in all M2 cytokines after placing M1 and M1 macrophages with inhibited STAT3 and STAT6 (M3 STAT3/6 phenotype) in the pro-tumor environment (constructed by data from ). AF – ascitic fluid.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques: Construct

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: The CD80/CD206 ratio in M1 macrophages cultured with AF and in M1 macrophages with inhibited STAT3 and STAT6 (M3 phenotype) cultured with AF.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques: Cell Culture

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Effect of M0, M1, and M3 STAT3/6 macrophages on the number of tumor cells. Experiment was performed in 5 replicates. w/o macs – without macrophages. Significance of differences from the M0 phenotype: * p<0.05; ** p<0.01

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques:

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Effects of M0, M1, and M3 STAT3/6 macrophages on cytokine microenvironment of tumor cells.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques:

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Changes in the ratio of mean% changes in all PI cytokines to mean% changes in all AI cytokines after addition of macrophages with M0, M1, and M3 STAT3/6 phenotypes (constructed by data from ). The ratio of PI-to-AI cytokines in the macrophage-free tumor microenvironment was taken as 1.0. A decrease in the ratio to below 1.0 means a shift of the microenvironment phenotype towards the anti-inflammatory phenotype, whereas an increase above 1.0 means a shift towards the pro-inflammatory phenotype. AI – anti-inflammatory; PI – pro-inflammatory.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques: Construct

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Effect of M3 SMAD3 , M3 STAT3/6+SMAD3 , and M3 STAT3/6 macrophages on the number of tumor cells. Experiment was performed in 5 replicates. w/o macs – without macrophages. Significance of differences from M3 SMAD3 macrophages: * p<0.05.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques:

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Effects of M3 STAT3/6 , M3 SMAD3 and M3 STAT3/6+SMAD3 macrophages on cytokine microenvironment of tumor cells.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques:

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Changes in the ratio of mean% changes in all PI cytokines to mean% changes in all AI cytokines after addition of macrophages with M3 STAT3/6 , M3 SMAD and M3 STAT3/6+SMAD phenotypes (constructed by data from ). The ratio of PI-to-AI cytokines in the macrophage-free tumor microenvironment was taken as 1.0. A decrease in the ratio to below 1.0 means a shift of the microenvironment phenotype towards the anti-inflammatory phenotype, whereas an increase above 1.0 means a shift towards the pro-inflammatory phenotype. AI – anti-inflammatory; PI – pro-inflammatory.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques: Construct

Journal: Medical Science Monitor Basic Research

Article Title: M3 Macrophages Stop Division of Tumor Cells In Vitro and Extend Survival of Mice with Ehrlich Ascites Carcinoma

doi: 10.12659/MSMBR.902285

Figure Lengend Snippet: Effect of injected M1 and M3 STAT3/6+SMAD macrophages and cisplatin on lifespan of mice, with EAC in Kaplan-Meier survival plots.

Article Snippet: We used the following reagents: Stat3 inhibitor (S3I204) (Axon Medchem, USA, cat# 2312), Stat6 inhibitor (As1517499) (Axon Medchem, USA, cat# 1992), IFN-γ (Invitrogen, USA, cat# PMC4034), LPS (Sigma-Aldrich, USA, cat# L3755), SMAD3 inhibitor (SIS3) (Calbiochem, USA, cat# 566405), FBS (Thermo Hyclone, UK, cat# SV30160.03), and cisplatin (TEVA, Israel).

Techniques: Injection

Journal: Cell Death & Disease

Article Title: CD45 + CD33 low CD11b dim myeloid-derived suppressor cells suppress CD8 + T cell activity via the IL-6/IL-8-arginase I axis in human gastric cancer

doi: 10.1038/s41419-018-0803-7

Figure Lengend Snippet: a Serum IL-6 and IL-8 concentration levels in GC patients compared to healthy donors. b Correlations between the proportions of peripheral blood CD45 + CD33 low CD11b dim myeloid cells or serum arginase I concentration and IL-6 or IL-8 serum concentrations in GC patients. c Arginase I expression and production in CD33 low CD11b dim myeloid cells exposed to recombinant IL-6 and/or IL-8, or 50% GC patient serum in the presence or absence of IL-6 and/or IL-8 neutralizing abs as analyzed by western blot and ELISA, respectively. d Arginase I expression in CD45 + CD33 low CD11b dim myeloid cells exposed to 50% GC patient serum in the presence or absence of FLLL32 (STAT3 phosphorylation inhibitor), Wortmannin (PI3K inhibitor), or SB203580 (MAPK inhibitor; top panel). AKT and p-AKT expression levels in CD45 + CD33 low CD11b dim myeloid cells exposed to IL-6 and/or IL-8 (middle panel), or 50% GC patient serum in the presence or absence of IL-6 and/or IL-8 neutralizing abs (bottom panels) as analyzed by western blot. * p < 0.05; ** p < 0.01, and n.s, p > 0.05. ARG I arginase I

Article Snippet: Drug inhibitors studies involved the STAT3 inhibitor FLLL32 (Medco Biosciences), PI3K inhibitor Wortmannin, and MAPK inhibitor SB203580 (Calbiochem).

Techniques: Concentration Assay, Expressing, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay