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: 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