stat3 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Cell Signaling Technology Inc stat3 p stat 3
    The role of TGase-1 in serum-stimulated nuclear accumulation of <t>Stat-3</t> and Stat-3 transcriptional activity. A , RPTC were cultured for 24 h in DMEM/F-12 with 5% FBS in the presence or absence of 100 μ m MDC and then stained with anti-Stat-3 antibody
    Stat3 P Stat 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stat3 p stat 3/product/Cell Signaling Technology Inc
    Average 99 stars, based on 38 article reviews
    Price from $9.99 to $1999.99
    stat3 p stat 3 - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc anti stat 3 antibodies
    Representative western blots and bar graphs of <t>STAT-3</t> protein expression by splenic mononuclear cells. STAT-3 expression is plotted as a ratio to PBS control (open bar). Treatment with ovine IL-6 increased STAT-3 (solid bar, n = 5, P
    Anti Stat 3 Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti stat 3 antibodies/product/Cell Signaling Technology Inc
    Average 94 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    anti stat 3 antibodies - by Bioz Stars, 2020-05
    94/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc phosphorylated stat 3 tyr705
    Representative western blots and bar graphs of <t>STAT-3</t> protein expression by splenic mononuclear cells. STAT-3 expression is plotted as a ratio to PBS control (open bar). Treatment with ovine IL-6 increased STAT-3 (solid bar, n = 5, P
    Phosphorylated Stat 3 Tyr705, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated stat 3 tyr705/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphorylated stat 3 tyr705 - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    88
    Santa Cruz Biotechnology stat 3
    Representative western blots and bar graphs of <t>STAT-3</t> protein expression by splenic mononuclear cells. STAT-3 expression is plotted as a ratio to PBS control (open bar). Treatment with ovine IL-6 increased STAT-3 (solid bar, n = 5, P
    Stat 3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stat 3/product/Santa Cruz Biotechnology
    Average 88 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    stat 3 - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    Image Search Results


    The role of TGase-1 in serum-stimulated nuclear accumulation of Stat-3 and Stat-3 transcriptional activity. A , RPTC were cultured for 24 h in DMEM/F-12 with 5% FBS in the presence or absence of 100 μ m MDC and then stained with anti-Stat-3 antibody

    Journal: The Journal of Biological Chemistry

    Article Title: Transglutaminase-1 Regulates Renal Epithelial Cell Proliferation through Activation of Stat-3 *

    doi: 10.1074/jbc.M808396200

    Figure Lengend Snippet: The role of TGase-1 in serum-stimulated nuclear accumulation of Stat-3 and Stat-3 transcriptional activity. A , RPTC were cultured for 24 h in DMEM/F-12 with 5% FBS in the presence or absence of 100 μ m MDC and then stained with anti-Stat-3 antibody

    Article Snippet: Chemicals and Antibodies —Antibodies to phospho-Stat-3, Stat-3, phospho-Akt, and Akt were purchased from Cell Signaling Technology (Danvers, MA).

    Techniques: Activity Assay, Cell Culture, Staining

    TGase-1 coexpression enhances JAK2-dependent phosphorylation of Stat-3. RPTC were transiently transfected with plasmids encoding wild types of TGase-1, JAK2, or Stat-3, either alone or in different combinations as indicated. After 48 h, the cells were

    Journal: The Journal of Biological Chemistry

    Article Title: Transglutaminase-1 Regulates Renal Epithelial Cell Proliferation through Activation of Stat-3 *

    doi: 10.1074/jbc.M808396200

    Figure Lengend Snippet: TGase-1 coexpression enhances JAK2-dependent phosphorylation of Stat-3. RPTC were transiently transfected with plasmids encoding wild types of TGase-1, JAK2, or Stat-3, either alone or in different combinations as indicated. After 48 h, the cells were

    Article Snippet: Chemicals and Antibodies —Antibodies to phospho-Stat-3, Stat-3, phospho-Akt, and Akt were purchased from Cell Signaling Technology (Danvers, MA).

    Techniques: Transfection

    Effect of MDC on phosphorylation of Akt, and Stat-3 in RPTC. A , RPTC were serum-starved for 24 h and then stimulated with 5% FBS for 1 h ( A ) or 4 and 8 h ( B ) in the presence or absence of 100 μ m MDC. Cell lysates were analyzed by immunoblot

    Journal: The Journal of Biological Chemistry

    Article Title: Transglutaminase-1 Regulates Renal Epithelial Cell Proliferation through Activation of Stat-3 *

    doi: 10.1074/jbc.M808396200

    Figure Lengend Snippet: Effect of MDC on phosphorylation of Akt, and Stat-3 in RPTC. A , RPTC were serum-starved for 24 h and then stimulated with 5% FBS for 1 h ( A ) or 4 and 8 h ( B ) in the presence or absence of 100 μ m MDC. Cell lysates were analyzed by immunoblot

    Article Snippet: Chemicals and Antibodies —Antibodies to phospho-Stat-3, Stat-3, phospho-Akt, and Akt were purchased from Cell Signaling Technology (Danvers, MA).

    Techniques:

    Effect of MDC, AG490, and Stat-3 siRNA on cyclin D1 and cyclin E expression. A and B , RPTC were serum-starved for 24 h and then incubated with 5% FBS for an additional 24 h in the presence or absence of 100 μ m MDC ( A ) or 10 μ m AG490

    Journal: The Journal of Biological Chemistry

    Article Title: Transglutaminase-1 Regulates Renal Epithelial Cell Proliferation through Activation of Stat-3 *

    doi: 10.1074/jbc.M808396200

    Figure Lengend Snippet: Effect of MDC, AG490, and Stat-3 siRNA on cyclin D1 and cyclin E expression. A and B , RPTC were serum-starved for 24 h and then incubated with 5% FBS for an additional 24 h in the presence or absence of 100 μ m MDC ( A ) or 10 μ m AG490

    Article Snippet: Chemicals and Antibodies —Antibodies to phospho-Stat-3, Stat-3, phospho-Akt, and Akt were purchased from Cell Signaling Technology (Danvers, MA).

    Techniques: Expressing, Incubation

    Effect of the JAK/Stat-3 pathway inhibition on RPTC proliferation. RPTC were cultured for 24 h in the presence of 5% FBS and then treated with AG490 at the indicated concentrations for an additional 24 h ( A ) or 1 h ( B ). A , cell proliferation was assessed

    Journal: The Journal of Biological Chemistry

    Article Title: Transglutaminase-1 Regulates Renal Epithelial Cell Proliferation through Activation of Stat-3 *

    doi: 10.1074/jbc.M808396200

    Figure Lengend Snippet: Effect of the JAK/Stat-3 pathway inhibition on RPTC proliferation. RPTC were cultured for 24 h in the presence of 5% FBS and then treated with AG490 at the indicated concentrations for an additional 24 h ( A ) or 1 h ( B ). A , cell proliferation was assessed

    Article Snippet: Chemicals and Antibodies —Antibodies to phospho-Stat-3, Stat-3, phospho-Akt, and Akt were purchased from Cell Signaling Technology (Danvers, MA).

    Techniques: Inhibition, Cell Culture

    Absence of STAT-3 activation in preadipocytes and adipocytes. The db/db, ob/ob , WT, or 3T3L1 preadipocytes were incubated in the presence or absence of LPS (1 μg/ml) for 1 hour. Cells were lysed subsequently, and Western blot analysis for total

    Journal:

    Article Title: Leptin-Dependent Toll-Like Receptor Expression and Responsiveness in Preadipocytes and Adipocytes

    doi: 10.2353/ajpath.2007.060699

    Figure Lengend Snippet: Absence of STAT-3 activation in preadipocytes and adipocytes. The db/db, ob/ob , WT, or 3T3L1 preadipocytes were incubated in the presence or absence of LPS (1 μg/ml) for 1 hour. Cells were lysed subsequently, and Western blot analysis for total

    Article Snippet: Antibodies against Akt, phospho-Akt (Ser473), STAT-3, as well as STAT-3P (Tyr705) were purchased from Cell Signaling (Beverly, MA).

    Techniques: Activation Assay, Incubation, Western Blot

    CD40 KO mice have attenuated transcription factor activity in lung interstitial cells compared to the activity in WT mice after CLP. WT or CD40 KO mice were subjected to CLP or a sham operation. (A) Lung interstitial cells were harvested, and the whole-cell extract was assayed for NF-κB by EMSA. WT and CD40 KO mice exhibited no NF-κB activity at the baseline (lanes 1 and 2). WT mice exhibited significant induction of NF-κB after CLP, which was reduced in CD40 KO mice (lanes 3 and 7). The specificity of the DNA binding complexes was confirmed by specific competition with excess unlabeled oligonucleotide (lanes 3 and 4). The lower band was determined to be an NF-κB-containing complex by supershift analysis with either anti-p65 or anti-p50 antibodies (compare lane 3 with lanes 5 and 6). All lanes contained pooled samples from two or three mice, and samples were normalized for protein. (B) Nuclear extracts from lung interstitial cells were assayed for the p65 subunit of NF-kB by immunoblotting. WT mice showed upregulation of p65 nuclear translocation after CLP (lanes 1 and 2). This was attenuated approximately twofold in CD40 KO mice (lanes 2 and 4). (C) Whole-cell extracts from lung interstitial cells were assayed for C/EBPβ by immunoblotting. WT mice exhibited a loss of C/EBPβ after CLP compared to the amount of C/EBPβ in controls that were not operated on (lanes 1 and 2). In contrast, there was little change in C/EBPβ in CD40 KO mice after CLP (lanes 3 and 4). (D) Lung interstitial cells were assayed for phosphorylated and total STAT-3 by immunoblotting. CD40 KO mice had attenuated STAT-3 phosphorylation in lung interstitial cells compared to the phosphorylation in WT mice after CLP (upper band) (compare lanes 3 and 4), and there was little difference in the total STAT-3 (lower band). There was no difference in the STAT-3 levels in sham-operated controls (lanes 1 and 2). Similar results were obtained with BALF cells (lanes 5 and 6). Finally, similar results were obtained with liver homogenates (lanes 7 and 8). Furthermore, the appearance of a nonspecific band (NS) provided an additional control for protein loading. In all cases, lanes were normalized for total protein. Lung interstitial cells from three to five mice were pooled.

    Journal: Infection and Immunity

    Article Title: CD40 Contributes to Lethality in Acute Sepsis: In Vivo Role for CD40 in Innate Immunity

    doi: 10.1128/IAI.71.6.3521-3528.2003

    Figure Lengend Snippet: CD40 KO mice have attenuated transcription factor activity in lung interstitial cells compared to the activity in WT mice after CLP. WT or CD40 KO mice were subjected to CLP or a sham operation. (A) Lung interstitial cells were harvested, and the whole-cell extract was assayed for NF-κB by EMSA. WT and CD40 KO mice exhibited no NF-κB activity at the baseline (lanes 1 and 2). WT mice exhibited significant induction of NF-κB after CLP, which was reduced in CD40 KO mice (lanes 3 and 7). The specificity of the DNA binding complexes was confirmed by specific competition with excess unlabeled oligonucleotide (lanes 3 and 4). The lower band was determined to be an NF-κB-containing complex by supershift analysis with either anti-p65 or anti-p50 antibodies (compare lane 3 with lanes 5 and 6). All lanes contained pooled samples from two or three mice, and samples were normalized for protein. (B) Nuclear extracts from lung interstitial cells were assayed for the p65 subunit of NF-kB by immunoblotting. WT mice showed upregulation of p65 nuclear translocation after CLP (lanes 1 and 2). This was attenuated approximately twofold in CD40 KO mice (lanes 2 and 4). (C) Whole-cell extracts from lung interstitial cells were assayed for C/EBPβ by immunoblotting. WT mice exhibited a loss of C/EBPβ after CLP compared to the amount of C/EBPβ in controls that were not operated on (lanes 1 and 2). In contrast, there was little change in C/EBPβ in CD40 KO mice after CLP (lanes 3 and 4). (D) Lung interstitial cells were assayed for phosphorylated and total STAT-3 by immunoblotting. CD40 KO mice had attenuated STAT-3 phosphorylation in lung interstitial cells compared to the phosphorylation in WT mice after CLP (upper band) (compare lanes 3 and 4), and there was little difference in the total STAT-3 (lower band). There was no difference in the STAT-3 levels in sham-operated controls (lanes 1 and 2). Similar results were obtained with BALF cells (lanes 5 and 6). Finally, similar results were obtained with liver homogenates (lanes 7 and 8). Furthermore, the appearance of a nonspecific band (NS) provided an additional control for protein loading. In all cases, lanes were normalized for total protein. Lung interstitial cells from three to five mice were pooled.

    Article Snippet: Membranes were probed with anti-phospho-STAT-3 or anti-STAT-3 (Cell Signal Technology, Beverly, Mass.).

    Techniques: Mouse Assay, Activity Assay, Binding Assay, Translocation Assay

    Proposed model for extracellular/intracellular Gal-9 trans and cis association with Tim-3 in regulation of monocyte inflammatory cytokine expressions through TLR signaling. After binding pathogenic products, TLR signaling involves cross-linking of adapter proteins, such as MyD88 (1) and TRIF (2), activation of NF К B-dependent/-independent (IRF3) pathways, and phosphorylation of multiple signaling molecules, including STAT-1 and STAT-3. Phosphorylated STAT-1 activates transcription/translation of IL-12 (3); whereas STAT-3 phosphorylation activates IL-23 but inhibits IL-12 expressions (4). In the case of chronic infections (such as HCV and HIV), pathogens may up-regulate Tim-3 expression on M/M Φ and Gal-9 expression by other cells, facilitating Tim-3/Gal-9 trans - association and so impairing innate immune responses. More specifically, extracellular Gal-9 trans association with Tim-3 expressed on the surface of M/M Φ , triggers Tim-3-SH2 tyrosine (Y265) phosphorylation by interleukin inducible T cell kinase (ITK), delivering negative signaling to TLR-mediated STAT-1 phosphorylation (5), and positive signaling to TLR-induced STAT-3 activation (6), resulting in inhibition of IL-12 but promotion of IL-23 productions. Imbalanced IL-23/IL-12 favors the differentiation of T H 2, TH17, and Treg cells that contribute to the development of chronic infection and autoimmune disorders. In the case of acute infection (such as self-limited HCV), TLR signaling may activate Gal-9 and Tim-3 cis association within the same M/M Φ (7) and yield adequate inflammatory cytokine expressions. Moreover, intracellular Gal-9 expressions can modulate M/M Φ Tim-3 and IL-12/IL-23 gene transcriptions (8). Upon TLR stimulation, intracellular Gal-9 rapidly translocates into nuclei, binds to NF-IL6 but not NF К B, and triggers IL-1β and IFN-γ transcriptions 31 (9). Intracellular Gal-9 also inhibits STAT3 but not STAT-1 phosphorylation, leading to increased IL-12 and decreased IL-23 gene transcriptions. However, intracellular Gal-9 appears to reduce Tim-3 cell surface expression by two distinct mechanisms: cis association/internalization and transcriptional inhibition, through a STAT-3-independent but possibly T-bet-dependent pathway (10). Appropriate TLR signaling in innate immune cells may thus facilitate proper adaptive immune responses to successfully clear the acute infection. Based on this schematic model, therapeutic strategies manipulating Tim-3/Gal-9 trans and/or cis association might be of clinical benefit.

    Journal: PLoS ONE

    Article Title: Cis Association of Galectin-9 with Tim-3 Differentially Regulates IL-12/IL-23 Expressions in Monocytes via TLR Signaling

    doi: 10.1371/journal.pone.0072488

    Figure Lengend Snippet: Proposed model for extracellular/intracellular Gal-9 trans and cis association with Tim-3 in regulation of monocyte inflammatory cytokine expressions through TLR signaling. After binding pathogenic products, TLR signaling involves cross-linking of adapter proteins, such as MyD88 (1) and TRIF (2), activation of NF К B-dependent/-independent (IRF3) pathways, and phosphorylation of multiple signaling molecules, including STAT-1 and STAT-3. Phosphorylated STAT-1 activates transcription/translation of IL-12 (3); whereas STAT-3 phosphorylation activates IL-23 but inhibits IL-12 expressions (4). In the case of chronic infections (such as HCV and HIV), pathogens may up-regulate Tim-3 expression on M/M Φ and Gal-9 expression by other cells, facilitating Tim-3/Gal-9 trans - association and so impairing innate immune responses. More specifically, extracellular Gal-9 trans association with Tim-3 expressed on the surface of M/M Φ , triggers Tim-3-SH2 tyrosine (Y265) phosphorylation by interleukin inducible T cell kinase (ITK), delivering negative signaling to TLR-mediated STAT-1 phosphorylation (5), and positive signaling to TLR-induced STAT-3 activation (6), resulting in inhibition of IL-12 but promotion of IL-23 productions. Imbalanced IL-23/IL-12 favors the differentiation of T H 2, TH17, and Treg cells that contribute to the development of chronic infection and autoimmune disorders. In the case of acute infection (such as self-limited HCV), TLR signaling may activate Gal-9 and Tim-3 cis association within the same M/M Φ (7) and yield adequate inflammatory cytokine expressions. Moreover, intracellular Gal-9 expressions can modulate M/M Φ Tim-3 and IL-12/IL-23 gene transcriptions (8). Upon TLR stimulation, intracellular Gal-9 rapidly translocates into nuclei, binds to NF-IL6 but not NF К B, and triggers IL-1β and IFN-γ transcriptions 31 (9). Intracellular Gal-9 also inhibits STAT3 but not STAT-1 phosphorylation, leading to increased IL-12 and decreased IL-23 gene transcriptions. However, intracellular Gal-9 appears to reduce Tim-3 cell surface expression by two distinct mechanisms: cis association/internalization and transcriptional inhibition, through a STAT-3-independent but possibly T-bet-dependent pathway (10). Appropriate TLR signaling in innate immune cells may thus facilitate proper adaptive immune responses to successfully clear the acute infection. Based on this schematic model, therapeutic strategies manipulating Tim-3/Gal-9 trans and/or cis association might be of clinical benefit.

    Article Snippet: Following transfer to an Amersham Hybond-P membrane (GE Heathcare, Piscataway, NJ), the membrane was blocked and probed with Gal-9 (Novus Biologicals, Littleton CO), Tim-3 (R & D, Mineapdis, MN), phospho-STAT-1 (Tyr701), phospho-STAT-3 (Tyr705) or total STAT-1, STAT-3 antibody (Cell Signaling Technology, Inc, Danvers, MA) or β-actin (Santa Cruz) at 4° C overnight.

    Techniques: Binding Assay, Activation Assay, Expressing, Inhibition, Infection

    Gal-9 differentially regulates IL-12 and IL-23 gene transcriptions in THP-1 cells through STAT-3 signaling. A) Western blot detection of Gal-9 and phosphorylation of STAT-1 proteins in THP-1 cells transfected with Gal-9 plasmid or siRNA. THP-1 cells were transfected with either pBKCMV3-Gal-9 plasmid (Gal-9) or pBKCMV3 empty vector (Con), or Gal-9 silencing siRNA or control siRNA. After 24~48 h transfection, the cells were subjected to Western blot analysis of Gal-9 and pSTAT-1 proteins. β-actin or total STAT-1 served as loading control to normalize target gene levels. Data are shown as representative imaging (left) and corrected optimal densitometry (O.D.) values from three independent experiments for STAT-1 (right). NS = no significance. B) Western blot detection of pSTAT-3 protein in THP-1 cells transfected with Gal-9 plasmid in the presence or absence of TLR stimulations. THP-1 cells were transfected with either Gal-9 or Control plasmid for 24 h, stimulated with or without LPS/R848 for 6 h, followed by Western blot analysis of pSTAT-3 protein. Total STAT-3 served as loading control to normalize target gene levels. Representative imaging and corrected O.D. values from three independent experiments are shown. *P

    Journal: PLoS ONE

    Article Title: Cis Association of Galectin-9 with Tim-3 Differentially Regulates IL-12/IL-23 Expressions in Monocytes via TLR Signaling

    doi: 10.1371/journal.pone.0072488

    Figure Lengend Snippet: Gal-9 differentially regulates IL-12 and IL-23 gene transcriptions in THP-1 cells through STAT-3 signaling. A) Western blot detection of Gal-9 and phosphorylation of STAT-1 proteins in THP-1 cells transfected with Gal-9 plasmid or siRNA. THP-1 cells were transfected with either pBKCMV3-Gal-9 plasmid (Gal-9) or pBKCMV3 empty vector (Con), or Gal-9 silencing siRNA or control siRNA. After 24~48 h transfection, the cells were subjected to Western blot analysis of Gal-9 and pSTAT-1 proteins. β-actin or total STAT-1 served as loading control to normalize target gene levels. Data are shown as representative imaging (left) and corrected optimal densitometry (O.D.) values from three independent experiments for STAT-1 (right). NS = no significance. B) Western blot detection of pSTAT-3 protein in THP-1 cells transfected with Gal-9 plasmid in the presence or absence of TLR stimulations. THP-1 cells were transfected with either Gal-9 or Control plasmid for 24 h, stimulated with or without LPS/R848 for 6 h, followed by Western blot analysis of pSTAT-3 protein. Total STAT-3 served as loading control to normalize target gene levels. Representative imaging and corrected O.D. values from three independent experiments are shown. *P

    Article Snippet: Following transfer to an Amersham Hybond-P membrane (GE Heathcare, Piscataway, NJ), the membrane was blocked and probed with Gal-9 (Novus Biologicals, Littleton CO), Tim-3 (R & D, Mineapdis, MN), phospho-STAT-1 (Tyr701), phospho-STAT-3 (Tyr705) or total STAT-1, STAT-3 antibody (Cell Signaling Technology, Inc, Danvers, MA) or β-actin (Santa Cruz) at 4° C overnight.

    Techniques: Western Blot, Transfection, Plasmid Preparation, Imaging

    The cardioprotective effect of S1P was abolished in cardiomyocyte-specific STAT-3 knockout mice subjected to ischaemia–reperfusion. In isolated hearts from cardiac-specific STAT-3- knockout mice, S1P failed to protect against an ischaemia–reperfusion insult. ( n ≥ 6 for all groups, * p

    Journal: Cardiovascular Journal of Africa

    Article Title: Cardiac preconditioning with sphingosine-1-phosphate requires activation of signal transducer and activator of transcription-3

    doi: 10.5830/CVJA-2014-016

    Figure Lengend Snippet: The cardioprotective effect of S1P was abolished in cardiomyocyte-specific STAT-3 knockout mice subjected to ischaemia–reperfusion. In isolated hearts from cardiac-specific STAT-3- knockout mice, S1P failed to protect against an ischaemia–reperfusion insult. ( n ≥ 6 for all groups, * p

    Article Snippet: Western blot analysis Phosphorylated and total STAT-3 levels were analysed by SDS polyacrylamide gel electrophoresis with antibodies from Cell Signalling Technology.

    Techniques: Knock-Out, Mouse Assay, Isolation

    Preconditioning protocols. (A) Schematic diagram of isolated mouse hearts undergoing a preconditioning protocol with and without S1P pre-treatment. (B) Schematic diagram of isolated mouse hearts undergoing a preconditioning protocol with and without S1P pre-treatment. These protocols were repeated in the presence of the STAT-3 inhibitor AG490.

    Journal: Cardiovascular Journal of Africa

    Article Title: Cardiac preconditioning with sphingosine-1-phosphate requires activation of signal transducer and activator of transcription-3

    doi: 10.5830/CVJA-2014-016

    Figure Lengend Snippet: Preconditioning protocols. (A) Schematic diagram of isolated mouse hearts undergoing a preconditioning protocol with and without S1P pre-treatment. (B) Schematic diagram of isolated mouse hearts undergoing a preconditioning protocol with and without S1P pre-treatment. These protocols were repeated in the presence of the STAT-3 inhibitor AG490.

    Article Snippet: Western blot analysis Phosphorylated and total STAT-3 levels were analysed by SDS polyacrylamide gel electrophoresis with antibodies from Cell Signalling Technology.

    Techniques: Isolation

    S1P pre-treatment increased phosphorylation of nuclear and mitochondrial STAT-3. Representative Western blots demonstrating levels of phosphorylated-STAT-3/total STAT-3 in (A) the cytoplasm, (B) the nucleus, and (C) the mitochondria after seven minutes of S1P pre-treatment in isolated rat hearts ( n = 4 per group, * p

    Journal: Cardiovascular Journal of Africa

    Article Title: Cardiac preconditioning with sphingosine-1-phosphate requires activation of signal transducer and activator of transcription-3

    doi: 10.5830/CVJA-2014-016

    Figure Lengend Snippet: S1P pre-treatment increased phosphorylation of nuclear and mitochondrial STAT-3. Representative Western blots demonstrating levels of phosphorylated-STAT-3/total STAT-3 in (A) the cytoplasm, (B) the nucleus, and (C) the mitochondria after seven minutes of S1P pre-treatment in isolated rat hearts ( n = 4 per group, * p

    Article Snippet: Western blot analysis Phosphorylated and total STAT-3 levels were analysed by SDS polyacrylamide gel electrophoresis with antibodies from Cell Signalling Technology.

    Techniques: Western Blot, Isolation

    S1P conferred protection via STAT-3 in the Langendorff-perfused rat heart. Co-incubation of the STAT-3 inhibitor AG490 (100 nmol/l) with S1P abolished the infarct-sparing effect of S1P in isolated rat hearts [ n > 6 per group, * p

    Journal: Cardiovascular Journal of Africa

    Article Title: Cardiac preconditioning with sphingosine-1-phosphate requires activation of signal transducer and activator of transcription-3

    doi: 10.5830/CVJA-2014-016

    Figure Lengend Snippet: S1P conferred protection via STAT-3 in the Langendorff-perfused rat heart. Co-incubation of the STAT-3 inhibitor AG490 (100 nmol/l) with S1P abolished the infarct-sparing effect of S1P in isolated rat hearts [ n > 6 per group, * p

    Article Snippet: Western blot analysis Phosphorylated and total STAT-3 levels were analysed by SDS polyacrylamide gel electrophoresis with antibodies from Cell Signalling Technology.

    Techniques: Incubation, Isolation

    Inhibitory effects of oleandrin and odoroside A on Oct3/4, β-catenin, and MMP-9 through the downregulation of STAT-3 phosphorylation. ( A ) Phospho-STAT-3 and total STAT-3 protein levels were detected in the cell lysates of ECs, MDA-MB-231 cells and RT-MDA-MB-231 cells by Western blot analysis. ( B – E ) MDA-MB-231 and RT-R-MDA-MB-231 cells were treated with oleandrin (50 nM), odoroside A (100 nM) and AG490 (10 μM; a phospho-STAT-3 inhibitor) ( B – D ) for 24 h. After treatment, phospho-STAT-3 ( B ), OCT3/4 and β-catenin ( C ) were determined by Western blot analysis, and MMP-9 activity was determined ( D ). The values are expressed as the means ± SEM from three independent determinations. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Oleandrin and Its Derivative Odoroside A, Both Cardiac Glycosides, Exhibit Anticancer Effects by Inhibiting Invasion via Suppressing the STAT-3 Signaling Pathway

    doi: 10.3390/ijms19113350

    Figure Lengend Snippet: Inhibitory effects of oleandrin and odoroside A on Oct3/4, β-catenin, and MMP-9 through the downregulation of STAT-3 phosphorylation. ( A ) Phospho-STAT-3 and total STAT-3 protein levels were detected in the cell lysates of ECs, MDA-MB-231 cells and RT-MDA-MB-231 cells by Western blot analysis. ( B – E ) MDA-MB-231 and RT-R-MDA-MB-231 cells were treated with oleandrin (50 nM), odoroside A (100 nM) and AG490 (10 μM; a phospho-STAT-3 inhibitor) ( B – D ) for 24 h. After treatment, phospho-STAT-3 ( B ), OCT3/4 and β-catenin ( C ) were determined by Western blot analysis, and MMP-9 activity was determined ( D ). The values are expressed as the means ± SEM from three independent determinations. * p

    Article Snippet: The membranes were blocked with 5% nonfat milk in Tris-buffered saline containing 0.05% Tween-20 for 1 h at room temperature and then incubated with the following primary antibodies: anti-OCT3/4 (sc-9081, 1:1000, rabbit polyclonal IgG, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-β-catenin (sc-7199, 1:1000, rabbit polyclonal IgG, Santa Cruz Biotechnology), anti-phospho-STAT-3 (9131S 1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-STAT-3 (4904S 1:1000, Cell Signaling Technology), and anti-SHP-1 (sc-7289 1:1000, Santa Cruz Biotechnology).

    Techniques: Multiple Displacement Amplification, Western Blot, Activity Assay

    Anticancer effects of oleandrin and odoroside A in MDA-MB-231 and RT-MDA-MB-231 cells through the downregulation of phospho-STAT-3. MDA-MB-231 and RT-MDA-MB-231 cells were treated as described in Figure 5 , and then the cells were collected and added to ECs-Matrigel-coated insert wells. The cells were incubated overnight (for 16 h) at 37 °C, and then the cells that had invaded across the membrane were stained with DAPI and counted as described in Figure 3 (×200 field image). The values are expressed as the means ± SEM from three independent determinations. ** p

    Journal: International Journal of Molecular Sciences

    Article Title: Oleandrin and Its Derivative Odoroside A, Both Cardiac Glycosides, Exhibit Anticancer Effects by Inhibiting Invasion via Suppressing the STAT-3 Signaling Pathway

    doi: 10.3390/ijms19113350

    Figure Lengend Snippet: Anticancer effects of oleandrin and odoroside A in MDA-MB-231 and RT-MDA-MB-231 cells through the downregulation of phospho-STAT-3. MDA-MB-231 and RT-MDA-MB-231 cells were treated as described in Figure 5 , and then the cells were collected and added to ECs-Matrigel-coated insert wells. The cells were incubated overnight (for 16 h) at 37 °C, and then the cells that had invaded across the membrane were stained with DAPI and counted as described in Figure 3 (×200 field image). The values are expressed as the means ± SEM from three independent determinations. ** p

    Article Snippet: The membranes were blocked with 5% nonfat milk in Tris-buffered saline containing 0.05% Tween-20 for 1 h at room temperature and then incubated with the following primary antibodies: anti-OCT3/4 (sc-9081, 1:1000, rabbit polyclonal IgG, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-β-catenin (sc-7199, 1:1000, rabbit polyclonal IgG, Santa Cruz Biotechnology), anti-phospho-STAT-3 (9131S 1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-STAT-3 (4904S 1:1000, Cell Signaling Technology), and anti-SHP-1 (sc-7289 1:1000, Santa Cruz Biotechnology).

    Techniques: Multiple Displacement Amplification, Incubation, Staining

    Inhibition of OSM-induced STAT-3 activation in human solid tumor cell lines by CK2 inhibitors. (A-C) Cell lysates were immunoblotted with the indicated antibodies. (A) CH235 human astroglioma cells were pretreated with TBB (50μM) for 2 hours and then stimulated with different concentrations of human OSM (0.5 and 1 ng/mL) for 30 minutes. The densitometric ratios of p-Y-STAT-3 versus STAT-3 were calculated. The value of lane 4 was compared with that of lane 3 (control, no inhibition), the value of lane 6 was compared with that of lane 5, and the percentage of inhibition was determined. γ2A-JAK2 human fibrosarcoma cells (B) and MDA-MB-231 human breast cancer cells (C) were pretreated with TBB (50μM) or emodin (50μM) for 2 hours and then stimulated with 0.5 ng/mL of human OSM for 30 minutes. The densitometric ratios of P-Y-STAT-3 versus STAT-3 were calculated. The values of lanes 3 and 4 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined.

    Journal: Blood

    Article Title: A CK2-dependent mechanism for activation of the JAK-STAT signaling pathway

    doi: 10.1182/blood-2010-01-266320

    Figure Lengend Snippet: Inhibition of OSM-induced STAT-3 activation in human solid tumor cell lines by CK2 inhibitors. (A-C) Cell lysates were immunoblotted with the indicated antibodies. (A) CH235 human astroglioma cells were pretreated with TBB (50μM) for 2 hours and then stimulated with different concentrations of human OSM (0.5 and 1 ng/mL) for 30 minutes. The densitometric ratios of p-Y-STAT-3 versus STAT-3 were calculated. The value of lane 4 was compared with that of lane 3 (control, no inhibition), the value of lane 6 was compared with that of lane 5, and the percentage of inhibition was determined. γ2A-JAK2 human fibrosarcoma cells (B) and MDA-MB-231 human breast cancer cells (C) were pretreated with TBB (50μM) or emodin (50μM) for 2 hours and then stimulated with 0.5 ng/mL of human OSM for 30 minutes. The densitometric ratios of P-Y-STAT-3 versus STAT-3 were calculated. The values of lanes 3 and 4 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined.

    Article Snippet: Antibodies to p-Y-STAT-3, total STAT-3, p-Y-STAT-1, p-Y-STAT-5, total STAT-5, p-JAK1, p-ERK, ERK, and caspase 8 were from Cell Signaling Technology.

    Techniques: Inhibition, Activation Assay, Multiple Displacement Amplification

    CK2 is required for OSM-induced STAT activation and gene expression. (A-E) Cell lysates were immunoblotted with the indicated antibodies. (A) MEFs were transfected with nontarget (NT) siRNA (100nM), CK2α siRNA (100nM), CK2β siRNA (100nM), or CK2α (50nM) plus CK2β siRNA (50nM) for 48 hours, then stimulated with 1 ng/mL of OSM for 10 minutes. The densitometric ratios of p-Y-STAT-3 versus STAT-3 were calculated. The values shown in lanes 4, 6, and 8 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined. (B) MEFs were pretreated with 25-100μM TBB for 2 hours and then stimulated with 1 ng/mL of OSM for 10 minutes. The densitometric ratios of p-Y-STAT-3 versus STAT-3 were calculated. The values shown in lanes 3, 4, and 5 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined. (C) MEFs were pretreated with TBB (50μM) for 2 and 4 hours and then stimulated with 0.1 ng/mL of OSM for 30 minutes. (D) MEFs were transfected with HA-CK2α or CK2α-inhibitor–resistant constructs for 24 hours, pretreated with TBB (50μM) for 2 hours and then stimulated with 1 ng/mL of OSM for 30 minutes. (E) MEFs were pretreated with CK2 inhibitors for 2 hours and then treated with OSM (10 ng/mL) for 30 minutes. (F) MEFs were transfected with nontarget (NT) siRNA (100nM), CK2α siRNA (100nM), CK2β siRNA (100nM), or CK2α (50nM) plus CK2β siRNA (50nM) for 48 hours, then stimulated with 0.1 ng/mL of OSM for 30 minutes. RNA was prepared and analyzed by RT-PCR for expression of SOCS-3 and GAPDH . The densitometric ratios of SOCS-3 versus GAPDH were calculated. The values of lanes 4, 6, and 8 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined. (G) MEFs were pretreated with TBB (50μM) for 2 hours and then stimulated with different concentrations of OSM (0.1-0.5 ng/mL). Total mRNA was extracted and analyzed by RPA with probes specific to SOCS-3 and GAPDH (loading control). The densitometric ratios of SOCS-3 versus GAPDH were calculated. The value of lane 4 was compared with that of lane 3 (control, no inhibition), the value of lane 6 was compared with that of lane 5, and the percentage of inhibition was determined.

    Journal: Blood

    Article Title: A CK2-dependent mechanism for activation of the JAK-STAT signaling pathway

    doi: 10.1182/blood-2010-01-266320

    Figure Lengend Snippet: CK2 is required for OSM-induced STAT activation and gene expression. (A-E) Cell lysates were immunoblotted with the indicated antibodies. (A) MEFs were transfected with nontarget (NT) siRNA (100nM), CK2α siRNA (100nM), CK2β siRNA (100nM), or CK2α (50nM) plus CK2β siRNA (50nM) for 48 hours, then stimulated with 1 ng/mL of OSM for 10 minutes. The densitometric ratios of p-Y-STAT-3 versus STAT-3 were calculated. The values shown in lanes 4, 6, and 8 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined. (B) MEFs were pretreated with 25-100μM TBB for 2 hours and then stimulated with 1 ng/mL of OSM for 10 minutes. The densitometric ratios of p-Y-STAT-3 versus STAT-3 were calculated. The values shown in lanes 3, 4, and 5 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined. (C) MEFs were pretreated with TBB (50μM) for 2 and 4 hours and then stimulated with 0.1 ng/mL of OSM for 30 minutes. (D) MEFs were transfected with HA-CK2α or CK2α-inhibitor–resistant constructs for 24 hours, pretreated with TBB (50μM) for 2 hours and then stimulated with 1 ng/mL of OSM for 30 minutes. (E) MEFs were pretreated with CK2 inhibitors for 2 hours and then treated with OSM (10 ng/mL) for 30 minutes. (F) MEFs were transfected with nontarget (NT) siRNA (100nM), CK2α siRNA (100nM), CK2β siRNA (100nM), or CK2α (50nM) plus CK2β siRNA (50nM) for 48 hours, then stimulated with 0.1 ng/mL of OSM for 30 minutes. RNA was prepared and analyzed by RT-PCR for expression of SOCS-3 and GAPDH . The densitometric ratios of SOCS-3 versus GAPDH were calculated. The values of lanes 4, 6, and 8 were compared with that of lane 2 (control, no inhibition) and the percentage of inhibition was determined. (G) MEFs were pretreated with TBB (50μM) for 2 hours and then stimulated with different concentrations of OSM (0.1-0.5 ng/mL). Total mRNA was extracted and analyzed by RPA with probes specific to SOCS-3 and GAPDH (loading control). The densitometric ratios of SOCS-3 versus GAPDH were calculated. The value of lane 4 was compared with that of lane 3 (control, no inhibition), the value of lane 6 was compared with that of lane 5, and the percentage of inhibition was determined.

    Article Snippet: Antibodies to p-Y-STAT-3, total STAT-3, p-Y-STAT-1, p-Y-STAT-5, total STAT-5, p-JAK1, p-ERK, ERK, and caspase 8 were from Cell Signaling Technology.

    Techniques: Activation Assay, Expressing, Transfection, Inhibition, Construct, Reverse Transcription Polymerase Chain Reaction, Recombinase Polymerase Amplification

    Inhibitory effects of oleandrin and odoroside A on Oct3/4, β-catenin, and MMP-9 through the downregulation of STAT-3 phosphorylation. ( A ) Phospho-STAT-3 and total STAT-3 protein levels were detected in the cell lysates of ECs, MDA-MB-231 cells and RT-MDA-MB-231 cells by Western blot analysis. ( B – E ) MDA-MB-231 and RT-R-MDA-MB-231 cells were treated with oleandrin (50 nM), odoroside A (100 nM) and AG490 (10 μM; a phospho-STAT-3 inhibitor) ( B – D ) for 24 h. After treatment, phospho-STAT-3 ( B ), OCT3/4 and β-catenin ( C ) were determined by Western blot analysis, and MMP-9 activity was determined ( D ). The values are expressed as the means ± SEM from three independent determinations. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Oleandrin and Its Derivative Odoroside A, Both Cardiac Glycosides, Exhibit Anticancer Effects by Inhibiting Invasion via Suppressing the STAT-3 Signaling Pathway

    doi: 10.3390/ijms19113350

    Figure Lengend Snippet: Inhibitory effects of oleandrin and odoroside A on Oct3/4, β-catenin, and MMP-9 through the downregulation of STAT-3 phosphorylation. ( A ) Phospho-STAT-3 and total STAT-3 protein levels were detected in the cell lysates of ECs, MDA-MB-231 cells and RT-MDA-MB-231 cells by Western blot analysis. ( B – E ) MDA-MB-231 and RT-R-MDA-MB-231 cells were treated with oleandrin (50 nM), odoroside A (100 nM) and AG490 (10 μM; a phospho-STAT-3 inhibitor) ( B – D ) for 24 h. After treatment, phospho-STAT-3 ( B ), OCT3/4 and β-catenin ( C ) were determined by Western blot analysis, and MMP-9 activity was determined ( D ). The values are expressed as the means ± SEM from three independent determinations. * p

    Article Snippet: Antibodies against OCT3/4, β-catenin and SHP-1 were purchased from Santa Cruz Biotechnology (Dallas, TX, USA), and antibodies against phospho-STAT-3 and STAT-3 were obtained from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Multiple Displacement Amplification, Western Blot, Activity Assay

    Western blot analysis of signal transducer and transcription (STAT)-3 and p-STAT-3 expressions in lymphocytes isolated from mouse colon lamina propria (LP). (a) The expressions of STAT-3 and p-STAT-3 were statistically analysed relative to β-actin

    Journal: Clinical and Experimental Immunology

    Article Title: Oral treatment with SEW2871, a sphingosine-1-phosphate type 1 receptor agonist, ameliorates experimental colitis in interleukin-10 gene deficient mice

    doi: 10.1111/cei.12304

    Figure Lengend Snippet: Western blot analysis of signal transducer and transcription (STAT)-3 and p-STAT-3 expressions in lymphocytes isolated from mouse colon lamina propria (LP). (a) The expressions of STAT-3 and p-STAT-3 were statistically analysed relative to β-actin

    Article Snippet: The primary antibodies against STAT-3 and p-STAT-3 were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Western Blot, Isolation

    iPSCs are less sensitive to LIF withdrawal than ESCs. ESCs or iPSCs were plated at a concentration of 8000 cells/cm 2 , harvested after 4 days and analyzed for Nanog-GFP expression by flow cytometry. Cells were grown in KO SR ( A ), KO SR supplemented with 10% (v/v) Hyclone serum (HY) ( B ) or GMEM supplemented with HY ( C ) in presence or absence of LIF, as indicated. Numbers shown on the histograms are the mean of FL1 fluorescence intensity (MFI). One representative experiment out of four is shown. D ESCs or iPSCs were seeded at 8500 cells/cm 2 in KO SR in the presence or absence of LIF. Protein extracts were prepared after 4 days incubation and immunoblotting performed with the antibodies indicated. Following detection of pY705 Stat-3, blots were stripped and reprobed to detect total levels of Stat-3. E and F Cells were seeded at 8500 cells/cm 2 in KO SR plus 10% (v/v) HY ( E ) or GMEM supplemented with 10% (v/v) HY ( F ), in the presence or absence of LIF. After 48 h protein extracts were prepared and immunoblotting performed as indicated.

    Journal: PLoS ONE

    Article Title: Differential Coupling of Self-Renewal Signaling Pathways in Murine Induced Pluripotent Stem Cells

    doi: 10.1371/journal.pone.0030234

    Figure Lengend Snippet: iPSCs are less sensitive to LIF withdrawal than ESCs. ESCs or iPSCs were plated at a concentration of 8000 cells/cm 2 , harvested after 4 days and analyzed for Nanog-GFP expression by flow cytometry. Cells were grown in KO SR ( A ), KO SR supplemented with 10% (v/v) Hyclone serum (HY) ( B ) or GMEM supplemented with HY ( C ) in presence or absence of LIF, as indicated. Numbers shown on the histograms are the mean of FL1 fluorescence intensity (MFI). One representative experiment out of four is shown. D ESCs or iPSCs were seeded at 8500 cells/cm 2 in KO SR in the presence or absence of LIF. Protein extracts were prepared after 4 days incubation and immunoblotting performed with the antibodies indicated. Following detection of pY705 Stat-3, blots were stripped and reprobed to detect total levels of Stat-3. E and F Cells were seeded at 8500 cells/cm 2 in KO SR plus 10% (v/v) HY ( E ) or GMEM supplemented with 10% (v/v) HY ( F ), in the presence or absence of LIF. After 48 h protein extracts were prepared and immunoblotting performed as indicated.

    Article Snippet: Immunoblotting was carried out with the following primary rabbit polyclonal antibodies: 1∶1000 for rabbit polyclonal antibodies recognizing dual phosphorylation of Erk1 and 2 at Thr202/Tyr204 (anti-pErk, Cell Signaling Technology 9101), phospho-tyrosine 705 of Stat-3 (anti-pSTAT-3, CST 9131), phospho-serine 473 of Akt (anti-pAkt, CST 9271), phospho-serines 21 or 9 of Gsk-3 α/β (anti-pGsk-3 α/β, CST 9331), phospho-serines 235/236 of S6 (anti-pS6 CST 4856), phospho-serines 411/418 of p70 S6 kinase (anti- p70S6 kinase CST 9204), phospho-serine 2448 of mTOR (anti-pmTOR CST 2976), phospho-serine 15 of p53 (anti p53 CST 9284), anti-pan β-catenin (CST 9562), anti-pan Akt (CST 9272); 1∶2000 anti-pan Erk (panErk, Santa Cruz Biotechnology, sc-93), anti-pan STAT-3 (panStat-3, sc-482), anti-pan SHP-2 (sc 293), anti-pan mTOR (CST 2983) anti-pan p70S6 kinase (CST 9202) anti β-Actin (SIGMA A5316) and anti GAPDH (CST 2118).

    Techniques: Concentration Assay, Expressing, Flow Cytometry, Cytometry, Fluorescence, Incubation

    Representative western blots and bar graphs of STAT-3 protein expression by splenic mononuclear cells. STAT-3 expression is plotted as a ratio to PBS control (open bar). Treatment with ovine IL-6 increased STAT-3 (solid bar, n = 5, P

    Journal: Journal of comparative pathology

    Article Title: In-vitro Validation of Cytokine Neutralizing Antibodies by testing with Ovine Mononuclear Splenocytes

    doi: 10.1016/j.jcpa.2012.06.001

    Figure Lengend Snippet: Representative western blots and bar graphs of STAT-3 protein expression by splenic mononuclear cells. STAT-3 expression is plotted as a ratio to PBS control (open bar). Treatment with ovine IL-6 increased STAT-3 (solid bar, n = 5, P

    Article Snippet: The membranes were probed with anti-IL-1β (rabbit polyclonal antibody; Lifespan Biosciences, Seattle, Washington, USA), anti-IL-6 (mouse mAb; Millipore, Billerica, Massachusetts, USA), anti-NF-κB (rabbit polyclonal antibody; Abcam, Cambridge, Massachusetts, USA), and anti-STAT-3 antibodies (rabbit polyclonal antibody; Cell Signalling, Danvers, Massachusetts, USA) at dilutions of 1 in 10,000, 1 in 5,000, 1 in 500 and 1 in 2,000, respectively.

    Techniques: Western Blot, Expressing

    Effects of Ovine IL-1β and IL-6 Protein Stimulation and Inhibition of Protein Stimulation by IL-1β and IL-6 mAbs on NF-κβ and STAT-3 Expression

    Journal: Journal of comparative pathology

    Article Title: In-vitro Validation of Cytokine Neutralizing Antibodies by testing with Ovine Mononuclear Splenocytes

    doi: 10.1016/j.jcpa.2012.06.001

    Figure Lengend Snippet: Effects of Ovine IL-1β and IL-6 Protein Stimulation and Inhibition of Protein Stimulation by IL-1β and IL-6 mAbs on NF-κβ and STAT-3 Expression

    Article Snippet: The membranes were probed with anti-IL-1β (rabbit polyclonal antibody; Lifespan Biosciences, Seattle, Washington, USA), anti-IL-6 (mouse mAb; Millipore, Billerica, Massachusetts, USA), anti-NF-κB (rabbit polyclonal antibody; Abcam, Cambridge, Massachusetts, USA), and anti-STAT-3 antibodies (rabbit polyclonal antibody; Cell Signalling, Danvers, Massachusetts, USA) at dilutions of 1 in 10,000, 1 in 5,000, 1 in 500 and 1 in 2,000, respectively.

    Techniques: Inhibition, Expressing

    Activation of TPOR signal transduction by antibody in AML cells. ( A ) After treatment with various doses of antibody or 10 ng/mL of TPO for 1 h, the phosphorylation of STAT-3, AKT, and ERK was analyzed by Western blotting using anti–p-STAT-3, anti–p-AKT,

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Agonist antibody that induces human malignant cells to kill one another

    doi: 10.1073/pnas.1519079112

    Figure Lengend Snippet: Activation of TPOR signal transduction by antibody in AML cells. ( A ) After treatment with various doses of antibody or 10 ng/mL of TPO for 1 h, the phosphorylation of STAT-3, AKT, and ERK was analyzed by Western blotting using anti–p-STAT-3, anti–p-AKT,

    Article Snippet: The anti–STAT-3, anti–p-STAT-3 (Tyr-705), anti-AKT, anti–p-AKT (Thr-308), anti-ERK1/2, and anti–p-ERK1/2 (Thr-202/Tyr-204) antibodies were purchased from Cell Signaling.

    Techniques: Activation Assay, Transduction, Western Blot