Journal: Infection and Immunity

Article Title: STAT3 Promotes Schistosome-Induced Liver Injury by Inflammation, Oxidative Stress, Proliferation, and Apoptosis Signal Pathway

doi: 10.1128/iai.00309-20

Figure Lengend Snippet: Figure 3 JAK2/STAT3 signal pathway was activated in the liver of mice with S. 623

Article Snippet: After dissolution in 10% or 12% SDS-PAGE, 433 the total protein samples were transferred to nitrocellulose membranes for 2 h with 434 transfer buffer and blocked with 5% nonfat dried milk, which were dissolved in TBST 435 (20 mmol/L Tris–HCl, pH 7.6, 150 mmol/L NaCl and 0.02% Tween 20) for 1 h. 436 Membranes were incubated with primary antibodies for GAPDH (Cell Signaling 437 Technology-2118, 1:1500 dilution), α-SMA (abcam-ab5694,1:800), IL-6 438 (abcam-ab6672), p-JAK2 (Cell Signaling Technology-3776), JAK2 (Cell Signaling 439 Technology-3230), p-STAT3 (Cell Signaling Technology-9145), STAT3 (Cell 440 on D ecem ber 17, 2020 at 43709427 http://iai.asm .org/ D ow nloaded from 21 Signaling Technology-9139/4904), Bax (Santa Cruz Biotechnology-SC493), Bcl-2 441 (Santa Cruz Biotechnology-SC492), Sirt3 (Cell Signaling Technology-5490s), 442 Acetylated SOD2 (K68) (abcam-ab137037), SOD2 (abcam-ab13534) overnight at 443 4°C.

Techniques:

Journal: Infection and Immunity

Article Title: STAT3 Promotes Schistosome-Induced Liver Injury by Inflammation, Oxidative Stress, Proliferation, and Apoptosis Signal Pathway

doi: 10.1128/iai.00309-20

Figure Lengend Snippet: Figure 4 Liver p-STAT3 deficiency attenuates hepatic injury caused by S. 635

Article Snippet: After dissolution in 10% or 12% SDS-PAGE, 433 the total protein samples were transferred to nitrocellulose membranes for 2 h with 434 transfer buffer and blocked with 5% nonfat dried milk, which were dissolved in TBST 435 (20 mmol/L Tris–HCl, pH 7.6, 150 mmol/L NaCl and 0.02% Tween 20) for 1 h. 436 Membranes were incubated with primary antibodies for GAPDH (Cell Signaling 437 Technology-2118, 1:1500 dilution), α-SMA (abcam-ab5694,1:800), IL-6 438 (abcam-ab6672), p-JAK2 (Cell Signaling Technology-3776), JAK2 (Cell Signaling 439 Technology-3230), p-STAT3 (Cell Signaling Technology-9145), STAT3 (Cell 440 on D ecem ber 17, 2020 at 43709427 http://iai.asm .org/ D ow nloaded from 21 Signaling Technology-9139/4904), Bax (Santa Cruz Biotechnology-SC493), Bcl-2 441 (Santa Cruz Biotechnology-SC492), Sirt3 (Cell Signaling Technology-5490s), 442 Acetylated SOD2 (K68) (abcam-ab137037), SOD2 (abcam-ab13534) overnight at 443 4°C.

Techniques:

Journal: Infection and Immunity

Article Title: STAT3 Promotes Schistosome-Induced Liver Injury by Inflammation, Oxidative Stress, Proliferation, and Apoptosis Signal Pathway

doi: 10.1128/iai.00309-20

Figure Lengend Snippet: Figure 6 Schematic overview of JAK2/STAT3-mediated effects on S. japonicum 668

Article Snippet: After dissolution in 10% or 12% SDS-PAGE, 433 the total protein samples were transferred to nitrocellulose membranes for 2 h with 434 transfer buffer and blocked with 5% nonfat dried milk, which were dissolved in TBST 435 (20 mmol/L Tris–HCl, pH 7.6, 150 mmol/L NaCl and 0.02% Tween 20) for 1 h. 436 Membranes were incubated with primary antibodies for GAPDH (Cell Signaling 437 Technology-2118, 1:1500 dilution), α-SMA (abcam-ab5694,1:800), IL-6 438 (abcam-ab6672), p-JAK2 (Cell Signaling Technology-3776), JAK2 (Cell Signaling 439 Technology-3230), p-STAT3 (Cell Signaling Technology-9145), STAT3 (Cell 440 on D ecem ber 17, 2020 at 43709427 http://iai.asm .org/ D ow nloaded from 21 Signaling Technology-9139/4904), Bax (Santa Cruz Biotechnology-SC493), Bcl-2 441 (Santa Cruz Biotechnology-SC492), Sirt3 (Cell Signaling Technology-5490s), 442 Acetylated SOD2 (K68) (abcam-ab137037), SOD2 (abcam-ab13534) overnight at 443 4°C.

Techniques:

Journal: Cell Death Discovery

Article Title: Interleukin-6 derived from cancer-associated fibroblasts attenuates the p53 response to doxorubicin in prostate cancer cells

doi: 10.1038/s41420-020-0272-5

Figure Lengend Snippet: a Levels of Tyr705-phosphorylated STAT3 (p-STAT3) and total STAT3 in LNCaP cells after treatment with 1 μM doxorubicin for 8 h in the presence or absence of IL-6 as shown by western blotting. Cells were pre-treated with IL-6 (0.1, 1 or 10 ng/mL) for 16 h ( N = 5). b Levels of Tyr705-phosphorylated STAT3 (p-STAT3) in 22Rv1 cells after treatment with 1 μM doxorubicin for 8 h in the presence or absence of IL-6 as shown by western blotting. Cells were pre-treated with 10 ng/mL IL-6 for 16 h ( N = 4). STAT3 phosphorylation at Tyr705 indicates STAT3 activation upon IL-6 treatment. The blots in a , b were cut into two pieces and probed with antibodies against p-STAT3 and GAPDH separately. The p-STAT3 blots were then stripped and blotted with an antibody against STAT3. c Survival of LNCaP cells as assessed by the WST assay after treatment with 1 μM doxorubicin for 48 h in the presence or absence of IL-6 and 1 μM JAK kinase inhibitor Ruxolitinib or 20 μM STAT3 inhibitor STA-21. Cells were pre-treated with IL-6 (0.1, 1 or 10 ng/mL) for 16 h. Cell survival is shown as fold change relative to cell survival in the absence of JAK/STAT3 inhibitor (S.E.M. is indicated by bars; N = 6). d Left panel: p53 levels in LNCaP cells after treatment with 1 μM doxorubicin for 8 h in the presence or absence of IL-6 and 1 μM JAK kinase inhibitor Ruxolitinib or 20 μM STAT3 inhibitor STA-21. Cells were pre-treated with IL-6 (10 ng/mL) for 16 h. The blot was cut into two pieces and probed with anti-p53 DO-1 and anti-GAPDH antibody separately. Samples to be compared were loaded on the same gel and transferred onto the same membrane. Right panel: quantification of the western blot data shown as ratio of p53 to GAPDH (S.E.M. is indicated by bars; N = 2). e STAT3 protein phosphorylation and total STAT3 protein levels in LNCaP cells after treatment with 1 μM doxorubicin for 8 h in the presence or absence of IL-6 and 1 μM JAK kinase inhibitor Ruxolitinib or 20 μM STAT3 inhibitor STA-21. Cells were pre-treated with 10 ng/mL IL-6 for 16 h ( N = 2). The blot was cut into two pieces and probed with antibodies against p-STAT3 and GAPDH separately. The p-STAT3 blot was then stripped and blotted with anti-STAT3 antibody.

Article Snippet: The inhibitors used were JAK inhibitors Ruxolitinib (INCB018424 Selleckchem, Rungsted, Denmark) and Pyridone 6 (Calbiochem) and STAT3 inhibitors STA-21 and Stattic (both from Selleckchem, Denmark).

Techniques: Western Blot, Phospho-proteomics, Activation Assay, WST Assay, Membrane

Journal: Cell Death Discovery

Article Title: Interleukin-6 derived from cancer-associated fibroblasts attenuates the p53 response to doxorubicin in prostate cancer cells

doi: 10.1038/s41420-020-0272-5

Figure Lengend Snippet: a Oncoprint from cBioportal of two selected prostate cancer studies showing genetic alteration profiles of IL-6R, STAT3, MDM2, and TP53. Each row represents the genetic alteration according to the figure legend with individual patient in the column. b mRNA Expression, RSEM (Batch normalized from Illumina HiSeq_RNASeqV2) of IL-6R, STAT3 or MDM2 from prostate adenocarcinoma (prad) of the TCGA PanCancer Atlas study based on patients that have no TP53 alterations or putative driver TP53 mutations (missense or truncating). N = 414 no alterations, and N = 56 putative driver mutations. Mann–Whitney test, for IL-6R ** p = 0.003, STAT3 p = 0.47 and MDM2 *** p < 0.0001. c Scatter plots of mRNA Expression, RSEM (Batch normalized from Illumina HiSeq_RNASeqV2) of IL-6R, JAK1, JAK2, STAT3 vs MDM2 in prostate adenocarcinoma patients of the TCGA PanCancer Atlas study that have no alterations in TP53. Indicated r and p values are analysis by Spearman correlation. N = 414 in all plots. d Scatter plots of mRNA expression, RSEM (Batch normalized from Illumina HiSeq_RNASeqV2) of IL-6R, JAK1, JAK2, STAT3 vs MDM2 in prostate adenocarcinoma patients of the TCGA PanCancer Atlas study that have putative driver alterations (missense and truncating) in TP53. Indicated r and p values are analysis by Spearman correlation. N = 56 in all plots, except the correlation with JAK2 N = 55. e Survival of patients with metastatic prostate adenocarcinoma from the study by Abida et al. . N is indicated in the figure. Black dot indicates censored patient. Left panel; patients with no TP53 alterations were grouped as having IL-6R alterations (amplification ( N = 12), mRNA high ( N = 1) missense mutation ( N = 1)) or having no IL-6R alterations. Log-rank (Mantel–Cox) test p = 0.09 or Gehan–Breslow–Wilcoxon test ** p = 0.01. Right panel; patients with putative driver TP53 mutations (missense, truncating, inframe) were grouped as having IL-6R alterations (amplification ( N = 4), mRNA high ( N = 3)) or having no IL-6R alterations. Log-rank (Mantel–Cox) test p = 0.5 or Gehan–Breslow–Wilcoxon test p = 0.9.

Article Snippet: The inhibitors used were JAK inhibitors Ruxolitinib (INCB018424 Selleckchem, Rungsted, Denmark) and Pyridone 6 (Calbiochem) and STAT3 inhibitors STA-21 and Stattic (both from Selleckchem, Denmark).

Techniques: Expressing, MANN-WHITNEY, Amplification, Mutagenesis

Journal: Cell Death Discovery

Article Title: Interleukin-6 derived from cancer-associated fibroblasts attenuates the p53 response to doxorubicin in prostate cancer cells

doi: 10.1038/s41420-020-0272-5

Figure Lengend Snippet: CAFs secrete IL-6, which binds to IL-6 receptors (IL-6R) expressed on cancer cells. IL-6 binding to its receptor leads to activation of JAK kinase and STAT3, respectively. STAT3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding. This stimulates Mdm2-mediated p53 ubiquitination and p53 degradation in the proteasome, thereby inhibiting upregulation of the p53 target Bax upon treatment with doxorubicin. As a result, prostate cancer cell survival is enhanced.

Article Snippet: The inhibitors used were JAK inhibitors Ruxolitinib (INCB018424 Selleckchem, Rungsted, Denmark) and Pyridone 6 (Calbiochem) and STAT3 inhibitors STA-21 and Stattic (both from Selleckchem, Denmark).

Techniques: Binding Assay, Activation Assay, Phospho-proteomics, Translocation Assay, Ubiquitin Proteomics

Journal: Scientific Reports

Article Title: WP1066 Sensitizes Oral Squamous Cell Carcinoma Cells to Cisplatin by Targeting STAT3/miR-21 axis

doi: 10.1038/srep07461

Figure Lengend Snippet: (a) In DDP resistant OSCC tumor tissues, STAT3 was highly expressed in the cytoplasm and nucleus of tumor cells than in DDP sensitive OSCC tissues ( P < 0.05). (B) In DDP resistant OSCC tissues, ISH staining reveals that miR-21 (red) has a relatively high expression than in DDP sensitive OSCC tissues ( P < 0.05) (Bar = 100 μm).

Article Snippet: Paraffin-embedded tumor tissue samples were selected for ISH with Cy3-labeled miR-21 probe (Boster, China) and IHC for STAT3 (Cell signaling, USA) expression.

Techniques: Staining, Expressing

Journal: Scientific Reports

Article Title: WP1066 Sensitizes Oral Squamous Cell Carcinoma Cells to Cisplatin by Targeting STAT3/miR-21 axis

doi: 10.1038/srep07461

Figure Lengend Snippet: (a) MTT assay indicated that Tca8113/DDP and Tca8113 cells showed different sensitivity to DDP treatment. The cells were incubated in different doses of DDP (0.1, 0.2, 0.5, 1, 2, 4, 8, 12 and 16 μg/ml) for 48 h, and survival rate was determined by MTT assay. (b) QPCR showed that miR-21 was 3.7 folds higher in Tca8113/DDP cells than in Tca8113 cells ( P < 0.05). (c) Western blot assay indicated that STAT3 protein expression was 3 folds higher in Tca8113/DDP cells than in Tca8113 cells ( P < 0.05). Anti-GAPDH antibody was used as a loading control.

Article Snippet: Paraffin-embedded tumor tissue samples were selected for ISH with Cy3-labeled miR-21 probe (Boster, China) and IHC for STAT3 (Cell signaling, USA) expression.

Techniques: MTT Assay, Incubation, Western Blot, Expressing

Journal: Scientific Reports

Article Title: WP1066 Sensitizes Oral Squamous Cell Carcinoma Cells to Cisplatin by Targeting STAT3/miR-21 axis

doi: 10.1038/srep07461

Figure Lengend Snippet: ISH was used to determine the expression of miR-21 in treated OSCC cells; and Western blot was used to detect the expression of its target genes. (a) In Tca8113 and Tca8113/DDP cells, WP1066 combined with DDP treatment could inhibit MMP2/9, Bcl-2, mTOR, Ki67 expression, while in DDP + WP1066 treated group, Caspase-3 expression was upregulated. (b) In Tca8113 and Tca8113/DDP cells, WP1066 combined with DDP treatment could induce the expression of miR-21 (Red signal labeled by Cy3, cell nuclei was labeled by DAPI). IL-6 treatment could inhibit the expression of miR-21. (Bar = 20 μm) (c) In Tca8113 and Tca8113/DDP cells, DDP + WP1066 treatment could induce the expression of PTEN, TIMP3 and PDCD4. While IL-6 treatment could inhibit their expression in both cell lines. The gels were cropped, but those gels were run under the same experimental condition.

Article Snippet: Paraffin-embedded tumor tissue samples were selected for ISH with Cy3-labeled miR-21 probe (Boster, China) and IHC for STAT3 (Cell signaling, USA) expression.

Techniques: Expressing, Western Blot, Labeling

Journal: Scientific Reports

Article Title: WP1066 Sensitizes Oral Squamous Cell Carcinoma Cells to Cisplatin by Targeting STAT3/miR-21 axis

doi: 10.1038/srep07461

Figure Lengend Snippet: (a) The growth curves of blank control, DMSO, WP1066-treated, DDP, and DDP + WP1066 treated Tca8113/DDP xenograft tumors. The growth rate in WP1066, DDP and DDP + WP1066 group were inhibited compared to control and DMSO groups. (b) ISH showed that, in WP1006, WP1066 and DDP treated Tca8113/DDP tumors, miR-21 expression was inhibited (Red signal labeled by Cy3, cell nuclei was labeled by DAPI). (c) TUNEL assay showed an induced apoptotic nucleus (green) in WP1066 combined DDP-treated Tca8113/DDP tumors. (d) IHC staining showed the upregulation of PTEN, TIMP-3, and PDCD4 in WP1066-treated and DDP + WP1066 treated group.(e) IHC assay was used to determine the expression of tumor growth related proteins in Tca8113/DDP xenograft tumors. Downregulation of mTOR, Ki67, Bcl-2, and MMP-2 was observed in WP1066 and WP1066 combined DDP-treated group. In combined treated group Caspase-3 expression was upregulated. (Bar = 100 μm).

Article Snippet: Paraffin-embedded tumor tissue samples were selected for ISH with Cy3-labeled miR-21 probe (Boster, China) and IHC for STAT3 (Cell signaling, USA) expression.

Techniques: Expressing, Labeling, TUNEL Assay, Immunohistochemistry

Journal: Nutrients

Article Title: Resveratrol Interferes with IL1-β-Induced Pro-Inflammatory Paracrine Interaction between Primary Chondrocytes and Macrophages

doi: 10.3390/nu8050280

Figure Lengend Snippet: Effects of a NF-κB inhibitor on the levels of p-NF-κB and p-STAT3 and on the levels of secreted IL-6, IL-8, Rantes and MCP-1 in HcH, MΦ cultures or in HcH/MΦ coculture. ( A ) Effect of a NF-κB inhibitor on the level of p-NF-κB (Western blot) and on the levels (ELISA) of IL-6, IL-8, Rantes, MCP-1 in HcH cultures and the effect of IL-1β treatment. Representative data from one of three experiments are shown; ( B ) Levels of p-STAT3 were determined by Western blotting) and the levels of IL-6 and IL-8 were determined by ELISA assay, in MΦ cultures. Effect of treatment with IL-1β, IL-6, aIL-6 or STA21. Co, control. Representative data from one of three experiments are shown; ( C ) Expression of p-STAT3 (Western blotting) and on the level (ELISA) of IL-6, IL-8, Rantes, MCP-1 in MΦ cocultures. Effect of IL-1β and NF-κB inhibitor. β-Tubulin: protein loading control, Pg/mL: picogram/mL. Representative data from one of three experiments are shown.

Article Snippet: The crucial role of IL-6 in our model was evaluated by using anti-IL-6 (from Merck Chimie SAS 201, Rue Carnot, Fontenay sous Bois, Île-de-France 94126, France), p65 antibody S536 (anti p-NF-κB) and Y705 (anti p-STAT3 antibody) (from Abcam, 24 rue Louis Blanc, 75010 PARIS, France), the NF-κB phosphorylation inhibitors (BAY-11-7082) (from Merck Chimie SAS, 201, Rue Carnot, Fontenay sous Bois, Île-de-France 94126, France), and STA21 (STAT3 phosphorylation inhibitor) (from Medchemexpress Europe Box 128-521-1298 Stockholm, Sweden).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Control, Expressing