stat1  (Abcam)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    STAT STAT1alpha STAT3 STAT5A STAT5B Transcription Factor Assay Kit Colorimetric
    Description:

    Catalog Number:
    ab207228
    Price:
    None
    Buy from Supplier


    Structured Review

    Abcam stat1
    Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of <t>STAT1,</t> IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.

    https://www.bioz.com/result/stat1/product/Abcam
    Average 99 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    stat1 - by Bioz Stars, 2020-11
    99/100 stars

    Images

    1) Product Images from "Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer"

    Article Title: Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer

    Journal: Oncoimmunology

    doi: 10.1080/2162402X.2017.1288330

    Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of STAT1, IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.
    Figure Legend Snippet: Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of STAT1, IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.

    Techniques Used: Expressing, Activation Assay, CTL Assay

    2) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    3) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    4) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    5) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    6) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    7) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    8) Product Images from "Short interfering RNA against STAT1 attenuates cisplatin-induced ototoxicity in the rat by suppressing inflammation"

    Article Title: Short interfering RNA against STAT1 attenuates cisplatin-induced ototoxicity in the rat by suppressing inflammation

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2011.63

    Cisplatin-induced hearing loss in rats is dependent on STAT1 activation. ( a ) ABR thresholds were recorded in Wistar rats treated with cisplatin (11 mg/kg, i.p.) for 72 h following 48 h trans-tympanic administration of scramble or STAT1 siRNA (0.9 μ g). ( b ) Scanning electron microscopic studies were performed on the cochleae. The representative image shows significant damage to OHCs (white arrows) by cisplatin, which is absent in the presence of STAT1 siRNA. ( c ) Quantitative analysis of the scanning electron micrographs. Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) indicate statistically significant difference from the scramble or the scramble+cisplatin treatment group ( P
    Figure Legend Snippet: Cisplatin-induced hearing loss in rats is dependent on STAT1 activation. ( a ) ABR thresholds were recorded in Wistar rats treated with cisplatin (11 mg/kg, i.p.) for 72 h following 48 h trans-tympanic administration of scramble or STAT1 siRNA (0.9 μ g). ( b ) Scanning electron microscopic studies were performed on the cochleae. The representative image shows significant damage to OHCs (white arrows) by cisplatin, which is absent in the presence of STAT1 siRNA. ( c ) Quantitative analysis of the scanning electron micrographs. Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) indicate statistically significant difference from the scramble or the scramble+cisplatin treatment group ( P

    Techniques Used: Activation Assay

    Cisplatin increases the expression and transcription of pro-inflammatory mediators through the STAT1-dependent pathway. ( a – c ) UB/OC-1 cells were transfected with scramble or STAT1 siRNA for 48 h, followed by cisplatin (2.5 μ M) for 24 h. Cell lysates were then used for western blotting studies to determine the levels of TNF- α ( a ), iNOS ( b ) and COX2 ( c ). β -Actin levels were used for normalization. ( d ) mRNA levels of iNOS, COX2 and TNF- α were determined by real-time RT-PCR in UB/OC-1 cultures treated as described in panels a – c . Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) denote statistically significant difference from the scramble- and the scramble+cisplatin-treated group, respectively ( P
    Figure Legend Snippet: Cisplatin increases the expression and transcription of pro-inflammatory mediators through the STAT1-dependent pathway. ( a – c ) UB/OC-1 cells were transfected with scramble or STAT1 siRNA for 48 h, followed by cisplatin (2.5 μ M) for 24 h. Cell lysates were then used for western blotting studies to determine the levels of TNF- α ( a ), iNOS ( b ) and COX2 ( c ). β -Actin levels were used for normalization. ( d ) mRNA levels of iNOS, COX2 and TNF- α were determined by real-time RT-PCR in UB/OC-1 cultures treated as described in panels a – c . Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) denote statistically significant difference from the scramble- and the scramble+cisplatin-treated group, respectively ( P

    Techniques Used: Expressing, Transfection, Western Blot, Quantitative RT-PCR

    ROS are essential for cisplatin-mediated STAT1 phosphorylation. ( a ) UB/OC-1 cells were pre-treated with vehicle or DPI (10 μ M), followed by cisplatin treatment for 45 min. STAT1 activation was determined by western blotting. H 2 O 2 (100 μ M) treatment served as the positive control for ROS generation. ( b ) Cells were transfected with scramble or NOX3 siRNA (5 nM) for 48 h, the culture media was replaced with fresh media and cells were exposed to vehicle or cisplatin (2.5 μ M) for 45 min. NOX3 siRNA blunted the activation of STAT1 by cisplatin without altering basal STAT1 levels. ( c ) UB/OC-1 cells were transfected with either scramble or NOX3 siRNA (5 nM) for 48 h, followed by cisplatin (2.5 μ M) treatment for another 24 h. NOX3 mRNA, measured by real-time RT-PCR, showed a 2.7±0.2-fold increase by cisplatin, which was reduced to 0.3±0.1-fold in the NOX3 siRNA+cisplatin group. NOX3 siRNA added alone reduced NOX3 mRNA to 0.3±0.1-fold. ( d ) ROS generation was measured in UB/OC-1 cells transfected with either scramble or NOX3 siRNA (5 nM) for 48 h, followed by treatment with cisplatin (2.5 μ M) for 15 min. Cells were then incubated with 5 μ M H 2 DCFDA dye for 15 min and ROS generation (green fluorescence) was visualized by confocal microscopy. DIC, differential interference contrast. Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) denote statistically significant difference from the vehicle/scramble- and the scramble+cisplatin-treated group, respectively ( P
    Figure Legend Snippet: ROS are essential for cisplatin-mediated STAT1 phosphorylation. ( a ) UB/OC-1 cells were pre-treated with vehicle or DPI (10 μ M), followed by cisplatin treatment for 45 min. STAT1 activation was determined by western blotting. H 2 O 2 (100 μ M) treatment served as the positive control for ROS generation. ( b ) Cells were transfected with scramble or NOX3 siRNA (5 nM) for 48 h, the culture media was replaced with fresh media and cells were exposed to vehicle or cisplatin (2.5 μ M) for 45 min. NOX3 siRNA blunted the activation of STAT1 by cisplatin without altering basal STAT1 levels. ( c ) UB/OC-1 cells were transfected with either scramble or NOX3 siRNA (5 nM) for 48 h, followed by cisplatin (2.5 μ M) treatment for another 24 h. NOX3 mRNA, measured by real-time RT-PCR, showed a 2.7±0.2-fold increase by cisplatin, which was reduced to 0.3±0.1-fold in the NOX3 siRNA+cisplatin group. NOX3 siRNA added alone reduced NOX3 mRNA to 0.3±0.1-fold. ( d ) ROS generation was measured in UB/OC-1 cells transfected with either scramble or NOX3 siRNA (5 nM) for 48 h, followed by treatment with cisplatin (2.5 μ M) for 15 min. Cells were then incubated with 5 μ M H 2 DCFDA dye for 15 min and ROS generation (green fluorescence) was visualized by confocal microscopy. DIC, differential interference contrast. Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) denote statistically significant difference from the vehicle/scramble- and the scramble+cisplatin-treated group, respectively ( P

    Techniques Used: Activation Assay, Western Blot, Positive Control, Transfection, Quantitative RT-PCR, Incubation, Fluorescence, Confocal Microscopy

    STAT1 siRNA reduced the cisplatin-mediated apoptosis of UB/OC-1 cells. ( a ) UB/OC-1 cells were transfected with scramble or STAT1 siRNA for 48 h, followed by cisplatin (20 μ M) for an additional 24 h. Apoptosis was determined by measuring the percentage of Annexin-positive and Annexin plus propidium iodide-positive cells (lower right and upper right quadrant respectively) by flow cytometry. ( b ) Percentage of apoptotic cells for each treatment as determined in panel a and plotted as the mean±S.E.M. ( * P
    Figure Legend Snippet: STAT1 siRNA reduced the cisplatin-mediated apoptosis of UB/OC-1 cells. ( a ) UB/OC-1 cells were transfected with scramble or STAT1 siRNA for 48 h, followed by cisplatin (20 μ M) for an additional 24 h. Apoptosis was determined by measuring the percentage of Annexin-positive and Annexin plus propidium iodide-positive cells (lower right and upper right quadrant respectively) by flow cytometry. ( b ) Percentage of apoptotic cells for each treatment as determined in panel a and plotted as the mean±S.E.M. ( * P

    Techniques Used: Transfection, Flow Cytometry, Cytometry

    Cisplatin activates STAT1 in UB/OC-1 cells and in the rat cochlea. ( a ) UB/OC-1 cells were treated with 2.5 μ M cisplatin for different time periods and STAT1 activation was determined by western blotting of whole-cell lysates. Activation of STAT1 (p-STAT1/STAT1) is indicated below. ( b ) UB/OC-1 cells were transfected with scramble siRNA or STAT1 siRNA for 48 h before cisplatin treatment for 45 min. Cell lysates were prepared and used in western blot studies for Ser 727 p-STAT1. ( c ) UB/OC-1 cells were co-transfected with a plasmid vector encoding STAT1 luciferase, along with a scramble or STAT1 siRNA. This was followed 24 h later with vehicle or cisplatin (2.5 μ M) for 8 h. Lysates were prepared and used for determination of luciferase activity. Co-transfection of a plasmid expressing Renilla luciferase allows normalization of luciferase activity in each well. ( d ) Immunolabeling studies were performed on the cochlear sections isolated from rats treated with vehicle or cisplatin (11 mg/kg, i.p.) for 72 h following trans-tympanic administration of scramble or STAT1 siRNA (0.9 μ g). Ser 727 p-STAT1 immunolabeling is indicated by green fluorescence whereas cell nuclei are defined by DAPI staining as indicated. Increased immunofluorescence is observed in the OHC, SVA, SG cells and SL. The scale bar shown in the lower right panel measures 50 μ m. ( e ) Magnified view of the OHC from panel d . The arrows indicate the three rows of OHCs. The scale bar shown in the lower right panel measures 10 μ m. Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) denote statistically significant difference from the vehicle/scramble- and the scramble+cisplatin-treated group, respectively ( P
    Figure Legend Snippet: Cisplatin activates STAT1 in UB/OC-1 cells and in the rat cochlea. ( a ) UB/OC-1 cells were treated with 2.5 μ M cisplatin for different time periods and STAT1 activation was determined by western blotting of whole-cell lysates. Activation of STAT1 (p-STAT1/STAT1) is indicated below. ( b ) UB/OC-1 cells were transfected with scramble siRNA or STAT1 siRNA for 48 h before cisplatin treatment for 45 min. Cell lysates were prepared and used in western blot studies for Ser 727 p-STAT1. ( c ) UB/OC-1 cells were co-transfected with a plasmid vector encoding STAT1 luciferase, along with a scramble or STAT1 siRNA. This was followed 24 h later with vehicle or cisplatin (2.5 μ M) for 8 h. Lysates were prepared and used for determination of luciferase activity. Co-transfection of a plasmid expressing Renilla luciferase allows normalization of luciferase activity in each well. ( d ) Immunolabeling studies were performed on the cochlear sections isolated from rats treated with vehicle or cisplatin (11 mg/kg, i.p.) for 72 h following trans-tympanic administration of scramble or STAT1 siRNA (0.9 μ g). Ser 727 p-STAT1 immunolabeling is indicated by green fluorescence whereas cell nuclei are defined by DAPI staining as indicated. Increased immunofluorescence is observed in the OHC, SVA, SG cells and SL. The scale bar shown in the lower right panel measures 50 μ m. ( e ) Magnified view of the OHC from panel d . The arrows indicate the three rows of OHCs. The scale bar shown in the lower right panel measures 10 μ m. Data are presented as mean±S.E.M. The asterisks ( * ) and ( ** ) denote statistically significant difference from the vehicle/scramble- and the scramble+cisplatin-treated group, respectively ( P

    Techniques Used: Activation Assay, Western Blot, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Cotransfection, Expressing, Immunolabeling, Isolation, Fluorescence, Staining, Immunofluorescence

    Cisplatin increases inflammation in rat cochlea. ( a ) Rats were treated with scramble or STAT1 siRNA by trans-tympanic injections followed by vehicle or cisplatin (11 mg/kg, i.p.) 48 h later. The rats were killed 72 h following the administration of vehicle or cisplatin. The cochleae were excised and processed for immunohistochemistry. Mid-modiolar sections of the cochlea were co-labeled with CD14 and TNF- α antibodies, followed by fluorescein- (green) or TRITC (red)-labeled secondary antibodies. Cisplatin increased CD14 and TNF- α immunoreactivity in cochleae treated with scramble siRNA. However, the increases in immunolabeling were attenuated in cochlea pretreated with STAT1 siRNA. The merged panels (yellow) indicate colocalization of CD14 and TNF- α . The scale bars (right lower panel) indicate 50 μ m. ( b ) Magnified view of the OHCs presented in panel a . The arrows indicate three rows of OHCs. The scale bar indicates 10 μ m
    Figure Legend Snippet: Cisplatin increases inflammation in rat cochlea. ( a ) Rats were treated with scramble or STAT1 siRNA by trans-tympanic injections followed by vehicle or cisplatin (11 mg/kg, i.p.) 48 h later. The rats were killed 72 h following the administration of vehicle or cisplatin. The cochleae were excised and processed for immunohistochemistry. Mid-modiolar sections of the cochlea were co-labeled with CD14 and TNF- α antibodies, followed by fluorescein- (green) or TRITC (red)-labeled secondary antibodies. Cisplatin increased CD14 and TNF- α immunoreactivity in cochleae treated with scramble siRNA. However, the increases in immunolabeling were attenuated in cochlea pretreated with STAT1 siRNA. The merged panels (yellow) indicate colocalization of CD14 and TNF- α . The scale bars (right lower panel) indicate 50 μ m. ( b ) Magnified view of the OHCs presented in panel a . The arrows indicate three rows of OHCs. The scale bar indicates 10 μ m

    Techniques Used: Immunohistochemistry, Labeling, Immunolabeling

    9) Product Images from "Type I Interferon Induced Epigenetic Regulation of Macrophages Suppresses Innate and Adaptive Immunity in Acute Respiratory Viral Infection"

    Article Title: Type I Interferon Induced Epigenetic Regulation of Macrophages Suppresses Innate and Adaptive Immunity in Acute Respiratory Viral Infection

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1005338

    Setdb2 expression by Mϕ is dependent on JAK-STAT pathway and IRF7. (A-C) RT-PCR of Setdb2 in BM-Mϕ (A, B) and CD11b + cells (C) from IAV-infected lungs. (A) Untreated and IFN-I-stimulated BM-Mϕ were treated with a vehicle control or the JAK inhibitor tofacitinib at the time of stimulation. (B) Control and Stat1 -/- BM-Mϕ were stimulated with media or IFN-I for 5 hours. (A, B) Data are mean ± SEM relative to unstimulated cells; n = 3 independent experiments. (C) CD11b + cells were isolated from the lungs of control and Stat1 -/- mice inoculated with IAV (1 x 10 4 PFU). Data are mean ± SEM relative to control cells on day 4 post-infection; n = 8–12 mice. (D, E) RT-PCR of Irf3 (D) and Irf7 (E) in control BM-Mϕ stimulated with IFN-I for the indicated time course. Data are mean ± SEM relative to unstimulated cells; n = 2–3 independent experiments (F, G) RT-PCR of Setdb2 and Irf3 / Irf7 in BM-Mϕ treated with control, IRF3, or IRF7 siRNA and stimulated with IFN-I for 24 hours. Data are mean ± SEM relative to cells treated with control siRNA; n = 3 independent experiments. (H, I) ChIP analysis of STAT1 (H) and IRF7 (I) binding in the Setdb2 promoter in control and IFN-I-stimulated BM- Mϕ after 24 hours. Data are mean ± SEM relative to IgG control; n = 2–3 independent experiments. N.D.; not detected. * p
    Figure Legend Snippet: Setdb2 expression by Mϕ is dependent on JAK-STAT pathway and IRF7. (A-C) RT-PCR of Setdb2 in BM-Mϕ (A, B) and CD11b + cells (C) from IAV-infected lungs. (A) Untreated and IFN-I-stimulated BM-Mϕ were treated with a vehicle control or the JAK inhibitor tofacitinib at the time of stimulation. (B) Control and Stat1 -/- BM-Mϕ were stimulated with media or IFN-I for 5 hours. (A, B) Data are mean ± SEM relative to unstimulated cells; n = 3 independent experiments. (C) CD11b + cells were isolated from the lungs of control and Stat1 -/- mice inoculated with IAV (1 x 10 4 PFU). Data are mean ± SEM relative to control cells on day 4 post-infection; n = 8–12 mice. (D, E) RT-PCR of Irf3 (D) and Irf7 (E) in control BM-Mϕ stimulated with IFN-I for the indicated time course. Data are mean ± SEM relative to unstimulated cells; n = 2–3 independent experiments (F, G) RT-PCR of Setdb2 and Irf3 / Irf7 in BM-Mϕ treated with control, IRF3, or IRF7 siRNA and stimulated with IFN-I for 24 hours. Data are mean ± SEM relative to cells treated with control siRNA; n = 3 independent experiments. (H, I) ChIP analysis of STAT1 (H) and IRF7 (I) binding in the Setdb2 promoter in control and IFN-I-stimulated BM- Mϕ after 24 hours. Data are mean ± SEM relative to IgG control; n = 2–3 independent experiments. N.D.; not detected. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Infection, Isolation, Mouse Assay, Chromatin Immunoprecipitation, Binding Assay

    Setdb2 suppresses the expression of antiviral genes and cytokine production nu BM-Mϕ. Gene expression (A, C) and cytokine production (B) by control and Setdb2 -/- BM-Mϕ stimulated with IFN-I for 24 hours. Data represents the mean ± SEM of 3 independent experiments. (A) Comparison of antiviral and proinflammatory genes by RT-PCR. Values on scatter plot represent log 10 (Δ Ct ). The gray lines represent a 2-fold change in gene expression; upregulated genes (red dots), downregulated genes (green dots). (B) Concentration of IL-1β, IL-6, IL-10, IL-12p40, G-GSF, and TNF-α in the supernatants of control and Setdb2 -/- BM-Mϕ. (C) RT-PCR of Isg15 and Mx1 . Data were normalized to control BM-Mϕ. (D-G) Control and Setdb2 -/- BM-Mϕ were treated with a vehicle control, IFN-I (D-G) , or IFN-II (F, G) for 24 hours. Data are mean ± SEM relative to IgG control; n = 2–3 experiments (D, E) ChIP analysis of Setdb2 binding and H3K9 tri-methylation in the Isg15 (D) and Mx1 (E) promoter. (F, G) ChIP analysis of STAT1 (F) and IRF7 (G) binding in the Mx1 promoter. (H) Control and Setdb2 ff Lyz2 cre+ mice inoculated with a sublethal dose of IAV (1 x 10 4 PFU). ) RT-PCR of Rnasel , Pkr , Mx1 , and Isg15 in infected lungs on day 4 post-infection. Data are mean ± SEM (n = 6–14 mice) from 2–3 independent experiments. * p
    Figure Legend Snippet: Setdb2 suppresses the expression of antiviral genes and cytokine production nu BM-Mϕ. Gene expression (A, C) and cytokine production (B) by control and Setdb2 -/- BM-Mϕ stimulated with IFN-I for 24 hours. Data represents the mean ± SEM of 3 independent experiments. (A) Comparison of antiviral and proinflammatory genes by RT-PCR. Values on scatter plot represent log 10 (Δ Ct ). The gray lines represent a 2-fold change in gene expression; upregulated genes (red dots), downregulated genes (green dots). (B) Concentration of IL-1β, IL-6, IL-10, IL-12p40, G-GSF, and TNF-α in the supernatants of control and Setdb2 -/- BM-Mϕ. (C) RT-PCR of Isg15 and Mx1 . Data were normalized to control BM-Mϕ. (D-G) Control and Setdb2 -/- BM-Mϕ were treated with a vehicle control, IFN-I (D-G) , or IFN-II (F, G) for 24 hours. Data are mean ± SEM relative to IgG control; n = 2–3 experiments (D, E) ChIP analysis of Setdb2 binding and H3K9 tri-methylation in the Isg15 (D) and Mx1 (E) promoter. (F, G) ChIP analysis of STAT1 (F) and IRF7 (G) binding in the Mx1 promoter. (H) Control and Setdb2 ff Lyz2 cre+ mice inoculated with a sublethal dose of IAV (1 x 10 4 PFU). ) RT-PCR of Rnasel , Pkr , Mx1 , and Isg15 in infected lungs on day 4 post-infection. Data are mean ± SEM (n = 6–14 mice) from 2–3 independent experiments. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Concentration Assay, Chromatin Immunoprecipitation, Binding Assay, Methylation, Mouse Assay, Infection

    10) Product Images from "Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression"

    Article Title: Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095627

    rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p
    Figure Legend Snippet: rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p

    Techniques Used: Immunofluorescence, Western Blot, Flow Cytometry, MANN-WHITNEY

    Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.
    Figure Legend Snippet: Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.

    Techniques Used:

    11) Product Images from "Essential role of Pin1 via STAT3 signalling and mitochondria-dependent pathways in restenosis in type 2 diabetes"

    Article Title: Essential role of Pin1 via STAT3 signalling and mitochondria-dependent pathways in restenosis in type 2 diabetes

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.12082

    Pin1 modulates cell growth and cell cycle progression by interfering with the STAT3 signalling pathway. Vascular smooth muscle cells (VSMCs) were cultured and infected as described in Figure 3 . The levels of STAT3, pTyr705-STAT3, pSer727-STAT3, STAT1, pTyr701-STAT1 and pSer727-STAT1 were quantified by Western blot analysis ( A and C ). The levels of some STAT3 downstream targets (p16ink4a, p21waf1/cip1, p27kip1 and survivin) in VSMCs were further observed by Western blot ( E ). Tubulin served as an internal control. Bar graph shows relative densitometric values of Western blots ( B , D and F ). Data represent mean ± SEM of three independent experiments. * P
    Figure Legend Snippet: Pin1 modulates cell growth and cell cycle progression by interfering with the STAT3 signalling pathway. Vascular smooth muscle cells (VSMCs) were cultured and infected as described in Figure 3 . The levels of STAT3, pTyr705-STAT3, pSer727-STAT3, STAT1, pTyr701-STAT1 and pSer727-STAT1 were quantified by Western blot analysis ( A and C ). The levels of some STAT3 downstream targets (p16ink4a, p21waf1/cip1, p27kip1 and survivin) in VSMCs were further observed by Western blot ( E ). Tubulin served as an internal control. Bar graph shows relative densitometric values of Western blots ( B , D and F ). Data represent mean ± SEM of three independent experiments. * P

    Techniques Used: Cell Culture, Infection, Western Blot

    12) Product Images from "The Coronavirus Transmissible Gastroenteritis Virus Evades the Type I Interferon Response through IRE1α-Mediated Manipulation of the MicroRNA miR-30a-5p/SOCS1/3 Axis"

    Article Title: The Coronavirus Transmissible Gastroenteritis Virus Evades the Type I Interferon Response through IRE1α-Mediated Manipulation of the MicroRNA miR-30a-5p/SOCS1/3 Axis

    Journal: Journal of Virology

    doi: 10.1128/JVI.00728-18

    Increased expression of SOCS1 or SOCS3 dampens the IFN-I antiviral response and promotes TGEV replication. (A) Overexpression of SOCS1 and SOCS3 in ST cells. ST cells were transfected as indicated with pCAGGS-HA, pCAGGS-SOCS1, or pCAGGS-SOCS3 for 48 h, and the transient expression of SOCS1 and SOCS3 was confirmed by IFA with anti-HA staining. (B) SOCS1 or SOCS3 overexpression suppressed the activation of STAT1 by IFN-β and TGEV infection. ST cells were transfected as indicated with pCAGGS-HA, pCAGGS-SOCS1, or pCAGGS-SOCS3 for 24 h, followed by incubation with porcine IFN-β (100 ng/ml) or infection with TGEV (MOI = 1). Cells were collected for Western blotting of pSTAT1, STAT1, or β-actin after 24 h. P values represent the difference from the vector control. (C, D) SOCS1 or SOCS3 overexpression enhanced TGEV infection and undermined the anti-TGEV activity of IFN-β. ST cells were transfected as described in the legend to panel B, followed by incubation with porcine IFN-β (C) or DMEM (D) for 24 h. Then, cells were infected with TGEV at an MOI of 0.01; TGEV infection was determined at 24 hpi. (E) Knockdown of SOCS1 and SOCS3 by siRNAs in ST cells. ST cells were harvested for Western blotting of SOCS1 and SOCS3 expression at 48 h after transfection with 100 nM siSOCS1s, siSOCS3s, or scrambled control siRNA. (F) Enhancement of the anti-TGEV activity of IFN-β by knockdown of SOCS1 or SOCS3 in ST cells. ST cells were transfected with siSOCS1s, siSOCS3s, or the scrambled control siRNA for 24 h, followed by incubation with IFN-β for 24 h. Then, the cells were infected with TGEV (MOI = 0.01) and harvested for quantification of TGEV infection at 24 hpi. (G) Silencing of SOCS1 or SOCS3 boosted IFN-β signaling under IFN-β-stimulated conditions. ST cells were stimulated with IFN-β at 24 h after transfection with siSOCS1#1, siSOCS3#3, or the scrambled control siRNA, and cells were collected for RT-qPCR analysis of ISG15, OASL, or MxA expression relative to that of GAPDH after 24 h of stimulation. (H) Silencing of SOCS1 or SOCS3 decreased TGEV infection under IFN-β-unstimulated conditions. ST cells were transfected with siSOCS1#1, siSOCS3#3, or the scrambled control siRNA for 24 h. Then, cells were infected TGEV (MOI = 0.01) and harvested for quantification of TGEV infection at 24 hpi. (I) Silencing of SOCS1 or SOCS3 boosted IFN-β signaling under TGEV-infected conditions. ST cells were treated as described in the legend to panel H, and cells were collected for RT-qPCR analysis of ISG15, OASL, or MxA expression relative to that of GAPDH. *, P
    Figure Legend Snippet: Increased expression of SOCS1 or SOCS3 dampens the IFN-I antiviral response and promotes TGEV replication. (A) Overexpression of SOCS1 and SOCS3 in ST cells. ST cells were transfected as indicated with pCAGGS-HA, pCAGGS-SOCS1, or pCAGGS-SOCS3 for 48 h, and the transient expression of SOCS1 and SOCS3 was confirmed by IFA with anti-HA staining. (B) SOCS1 or SOCS3 overexpression suppressed the activation of STAT1 by IFN-β and TGEV infection. ST cells were transfected as indicated with pCAGGS-HA, pCAGGS-SOCS1, or pCAGGS-SOCS3 for 24 h, followed by incubation with porcine IFN-β (100 ng/ml) or infection with TGEV (MOI = 1). Cells were collected for Western blotting of pSTAT1, STAT1, or β-actin after 24 h. P values represent the difference from the vector control. (C, D) SOCS1 or SOCS3 overexpression enhanced TGEV infection and undermined the anti-TGEV activity of IFN-β. ST cells were transfected as described in the legend to panel B, followed by incubation with porcine IFN-β (C) or DMEM (D) for 24 h. Then, cells were infected with TGEV at an MOI of 0.01; TGEV infection was determined at 24 hpi. (E) Knockdown of SOCS1 and SOCS3 by siRNAs in ST cells. ST cells were harvested for Western blotting of SOCS1 and SOCS3 expression at 48 h after transfection with 100 nM siSOCS1s, siSOCS3s, or scrambled control siRNA. (F) Enhancement of the anti-TGEV activity of IFN-β by knockdown of SOCS1 or SOCS3 in ST cells. ST cells were transfected with siSOCS1s, siSOCS3s, or the scrambled control siRNA for 24 h, followed by incubation with IFN-β for 24 h. Then, the cells were infected with TGEV (MOI = 0.01) and harvested for quantification of TGEV infection at 24 hpi. (G) Silencing of SOCS1 or SOCS3 boosted IFN-β signaling under IFN-β-stimulated conditions. ST cells were stimulated with IFN-β at 24 h after transfection with siSOCS1#1, siSOCS3#3, or the scrambled control siRNA, and cells were collected for RT-qPCR analysis of ISG15, OASL, or MxA expression relative to that of GAPDH after 24 h of stimulation. (H) Silencing of SOCS1 or SOCS3 decreased TGEV infection under IFN-β-unstimulated conditions. ST cells were transfected with siSOCS1#1, siSOCS3#3, or the scrambled control siRNA for 24 h. Then, cells were infected TGEV (MOI = 0.01) and harvested for quantification of TGEV infection at 24 hpi. (I) Silencing of SOCS1 or SOCS3 boosted IFN-β signaling under TGEV-infected conditions. ST cells were treated as described in the legend to panel H, and cells were collected for RT-qPCR analysis of ISG15, OASL, or MxA expression relative to that of GAPDH. *, P

    Techniques Used: Expressing, Over Expression, Transfection, Immunofluorescence, Staining, Activation Assay, Infection, Incubation, Western Blot, Plasmid Preparation, Activity Assay, Quantitative RT-PCR

    miR-30a-5p enhances IFN-I antiviral signaling rather than IFN-I production. (A) miR-30a-5p did not manipulate IFN-β production. ST cells were transfected with 160 nM miR-30a-5p mimics or NC mimics for 24 h, followed by infection with TGEV (MOI = 0.01) for 24 h. The IFN-β levels in the supernatant were measured by ELISA. NS, not significant. (B, C) TGEV infection antagonized interferon signaling at the late stages of infection. ST cells were infected with TGEV at an MOI of 1, and then the samples were collected at different times for the quantification of ISG15, IFN-β, and miR-30a-5p expression (B) or the Western blotting of pSTAT1, STAT1, or β-actin (C). *, P
    Figure Legend Snippet: miR-30a-5p enhances IFN-I antiviral signaling rather than IFN-I production. (A) miR-30a-5p did not manipulate IFN-β production. ST cells were transfected with 160 nM miR-30a-5p mimics or NC mimics for 24 h, followed by infection with TGEV (MOI = 0.01) for 24 h. The IFN-β levels in the supernatant were measured by ELISA. NS, not significant. (B, C) TGEV infection antagonized interferon signaling at the late stages of infection. ST cells were infected with TGEV at an MOI of 1, and then the samples were collected at different times for the quantification of ISG15, IFN-β, and miR-30a-5p expression (B) or the Western blotting of pSTAT1, STAT1, or β-actin (C). *, P

    Techniques Used: Transfection, Infection, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

    IRE1α facilitates TGEV infection by modulating the miR-30a-5p/SOCS axis. (A, B) The blockage of STAT1 activation rescued the viral suppression of 4μ8c. ST cells were pretreated with 100 μM 4μ8c for 2 h or 100 μM 4μ8c for 2 h plus 10 μM fludarabine (Flud) for 24 h, followed by infection with TGEV (MOI = 1). Then, the TGEV titer (A) was measured at 24 hpi, and STAT1 and p-STAT1 were analyzed by Western blotting (B). (C, D) The blockage of STAT1 activation rescued the viral suppression of miR-30a-5p. ST cells were transfected with 160 nM NC mimics, miR-30a-5p mimics, or miR-30a-5p mimics plus stimulation with 10 μM fludarabine for 24 h, followed by infection with TGEV (MOI = 1). Next, the TGEV titer (C) was measured at 24 hpi, and STAT1 and p-STAT1 were analyzed by Western blotting (D). (E) Effect of 4μ8c and fludarabine on cell viability. ST cells were treated with 4μ8c, 4μ8c plus fludarabine, fludarabine only, or the carrier control (DMSO) as described above. Cell cytotoxicity was analyzed with a CCK-8 system as described in Materials and Methods. (F) Knockdown of IFNAR1 by siRNAs in ST cells. ST cells were harvested for RT-qPCR analysis of IFNAR1 expression at 48 h after transfection with 100 nM siRNAs or scrambled control siRNA (siCtrl). (G) Silencing of IFNAR1 dampened IFN-β signaling under IFN-β-stimulated conditions. ST cells were transfected with 100 nM siIFNAR1#1 or scrambled control siRNA for 24 h, followed by incubation with IFN-β for 24 h. Then, the cells were collected for RT-qPCR analysis of ISG15, OASL, or MxA expression relative to that of GAPDH. (H, I) Silencing of IFNAR1 abolished viral suppression (H) and enhancement of p-STAT1 (I) of miR-30a-5p. NC mimics or miR-30a-5p mimics (160 nM) were cotransfected with 100 nM siIFNAR1#1 or the scrambled control siRNA in ST cells for 24 h, followed by infection with TGEV (MOI = 1). The TGEV titer (H) and STAT1 and p-STAT1 levels (I) were measured at 24 hpi. *, P
    Figure Legend Snippet: IRE1α facilitates TGEV infection by modulating the miR-30a-5p/SOCS axis. (A, B) The blockage of STAT1 activation rescued the viral suppression of 4μ8c. ST cells were pretreated with 100 μM 4μ8c for 2 h or 100 μM 4μ8c for 2 h plus 10 μM fludarabine (Flud) for 24 h, followed by infection with TGEV (MOI = 1). Then, the TGEV titer (A) was measured at 24 hpi, and STAT1 and p-STAT1 were analyzed by Western blotting (B). (C, D) The blockage of STAT1 activation rescued the viral suppression of miR-30a-5p. ST cells were transfected with 160 nM NC mimics, miR-30a-5p mimics, or miR-30a-5p mimics plus stimulation with 10 μM fludarabine for 24 h, followed by infection with TGEV (MOI = 1). Next, the TGEV titer (C) was measured at 24 hpi, and STAT1 and p-STAT1 were analyzed by Western blotting (D). (E) Effect of 4μ8c and fludarabine on cell viability. ST cells were treated with 4μ8c, 4μ8c plus fludarabine, fludarabine only, or the carrier control (DMSO) as described above. Cell cytotoxicity was analyzed with a CCK-8 system as described in Materials and Methods. (F) Knockdown of IFNAR1 by siRNAs in ST cells. ST cells were harvested for RT-qPCR analysis of IFNAR1 expression at 48 h after transfection with 100 nM siRNAs or scrambled control siRNA (siCtrl). (G) Silencing of IFNAR1 dampened IFN-β signaling under IFN-β-stimulated conditions. ST cells were transfected with 100 nM siIFNAR1#1 or scrambled control siRNA for 24 h, followed by incubation with IFN-β for 24 h. Then, the cells were collected for RT-qPCR analysis of ISG15, OASL, or MxA expression relative to that of GAPDH. (H, I) Silencing of IFNAR1 abolished viral suppression (H) and enhancement of p-STAT1 (I) of miR-30a-5p. NC mimics or miR-30a-5p mimics (160 nM) were cotransfected with 100 nM siIFNAR1#1 or the scrambled control siRNA in ST cells for 24 h, followed by infection with TGEV (MOI = 1). The TGEV titer (H) and STAT1 and p-STAT1 levels (I) were measured at 24 hpi. *, P

    Techniques Used: Infection, Activation Assay, Western Blot, Transfection, CCK-8 Assay, Quantitative RT-PCR, Expressing, Incubation

    13) Product Images from "The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney"

    Article Title: The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1005734

    Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P
    Figure Legend Snippet: Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P

    Techniques Used: Expressing, Activation Assay, Staining

    Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (
    Figure Legend Snippet: Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (

    Techniques Used: Activity Assay, Expressing

    Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).
    Figure Legend Snippet: Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).

    Techniques Used: Activation Assay, Expressing, Microarray

    14) Product Images from "SARS Coronavirus Papain-Like Protease Inhibits the TLR7 Signaling Pathway through Removing Lys63-Linked Polyubiquitination of TRAF3 and TRAF6"

    Article Title: SARS Coronavirus Papain-Like Protease Inhibits the TLR7 Signaling Pathway through Removing Lys63-Linked Polyubiquitination of TRAF3 and TRAF6

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms17050678

    Inhibitory effect of SARS-CoV PLPro on TLR7 agonist-induced activation of type I IFN signaling. ISRE-driven luciferase reporter activity and the mRNA levels of PKR and IRF7 were determined 4 h post-IMQ treatment. ISRE-driven firefly luciferase activity was normalized by Renilla luciferase activity ( A ). Relative mRNA levels of PKR ( B ) and IRF7 ( C ) were normalized by GAPDH mRNA, presented as a relative ratio. In addition, the activated status of STAT1 was examined using Western blot with anti-phospho-STAT1 (Tyr701) antibodies ( D ). * p -Value
    Figure Legend Snippet: Inhibitory effect of SARS-CoV PLPro on TLR7 agonist-induced activation of type I IFN signaling. ISRE-driven luciferase reporter activity and the mRNA levels of PKR and IRF7 were determined 4 h post-IMQ treatment. ISRE-driven firefly luciferase activity was normalized by Renilla luciferase activity ( A ). Relative mRNA levels of PKR ( B ) and IRF7 ( C ) were normalized by GAPDH mRNA, presented as a relative ratio. In addition, the activated status of STAT1 was examined using Western blot with anti-phospho-STAT1 (Tyr701) antibodies ( D ). * p -Value

    Techniques Used: Activation Assay, Luciferase, Activity Assay, Western Blot

    15) Product Images from "Prolonged TNFα primes fibroblast-like synoviocytes in a gene-specific manner by altering chromatin"

    Article Title: Prolonged TNFα primes fibroblast-like synoviocytes in a gene-specific manner by altering chromatin

    Journal: Arthritis & rheumatology (Hoboken, N.J.)

    doi: 10.1002/art.38871

    STAT1 amplifier function of chronic TNFα in RA FLS RA FLS were cultured for 3 days in the presence or absence of TNFα (10 ng/ml), which was added on the first day of culture and was not replenished ( A, D ). ( A ), On day 3 the cells were stimulated with IFNβ (1,000 U/ml) or IFNγ (100 U/ml) for 10-60 minutes and STAT1 tyrosine phosphorylation (pY) was measured by immunoblotting. The expression of STAT1 protein ( B ) and mRNA ( C ), upon TNFα time course stimulation, was measured by immunoblotting and qPCR. Values are the mean ±SEM and were normalized relative to mRNA for GAPDH. *= p
    Figure Legend Snippet: STAT1 amplifier function of chronic TNFα in RA FLS RA FLS were cultured for 3 days in the presence or absence of TNFα (10 ng/ml), which was added on the first day of culture and was not replenished ( A, D ). ( A ), On day 3 the cells were stimulated with IFNβ (1,000 U/ml) or IFNγ (100 U/ml) for 10-60 minutes and STAT1 tyrosine phosphorylation (pY) was measured by immunoblotting. The expression of STAT1 protein ( B ) and mRNA ( C ), upon TNFα time course stimulation, was measured by immunoblotting and qPCR. Values are the mean ±SEM and were normalized relative to mRNA for GAPDH. *= p

    Techniques Used: Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    16) Product Images from "Age-Dependent and -Independent Effects of Perivascular Adipose Tissue and Its Paracrine Activities during Neointima Formation"

    Article Title: Age-Dependent and -Independent Effects of Perivascular Adipose Tissue and Its Paracrine Activities during Neointima Formation

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21010282

    Perivascular adipose tissue expression of negative regulators of STAT1 and NFκB. ( A , B ) Western blot analysis of protein inhibitor of activated STAT-1 (PIAS1), ( A , C ) suppressor of cytokine signaling-1 (SOCS1), ( D , E ) myeloid differentiation primary response-88 (MyD88), and ( D , F ) tumor suppressor p53 (p53) in visceral (VAT) and perivascular (PVAT) adipose tissue of adult ( n = 3–8) and middle-aged ( n = 3–8) mice. Results were normalized to GAPDH protein levels and are expressed as -fold change vs. findings obtained in VAT of adult mice. Individual data points and the mean ± SD are shown. # p
    Figure Legend Snippet: Perivascular adipose tissue expression of negative regulators of STAT1 and NFκB. ( A , B ) Western blot analysis of protein inhibitor of activated STAT-1 (PIAS1), ( A , C ) suppressor of cytokine signaling-1 (SOCS1), ( D , E ) myeloid differentiation primary response-88 (MyD88), and ( D , F ) tumor suppressor p53 (p53) in visceral (VAT) and perivascular (PVAT) adipose tissue of adult ( n = 3–8) and middle-aged ( n = 3–8) mice. Results were normalized to GAPDH protein levels and are expressed as -fold change vs. findings obtained in VAT of adult mice. Individual data points and the mean ± SD are shown. # p

    Techniques Used: Expressing, Western Blot, Mouse Assay

    17) Product Images from "The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney"

    Article Title: The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1005734

    Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P
    Figure Legend Snippet: Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P

    Techniques Used: Expressing, Activation Assay, Staining

    Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (
    Figure Legend Snippet: Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (

    Techniques Used: Activity Assay, Expressing

    Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).
    Figure Legend Snippet: Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).

    Techniques Used: Activation Assay, Expressing, Microarray

    18) Product Images from "TIR-domain-containing adapter-inducing interferon-β (TRIF) regulates Th17-mediated intestinal immunopathology in colitis"

    Article Title: TIR-domain-containing adapter-inducing interferon-β (TRIF) regulates Th17-mediated intestinal immunopathology in colitis

    Journal: Mucosal Immunology

    doi: 10.1038/mi.2014.67

    Interleukin (IL)-27p28-induced STAT1 activation regulates Th17-cell differentiation and plasticity. ( a ) Western blot analysis of transcription factors in wild-type (WT) naive T cells after co-culturing with MΦs in the presence of cecal bacterial antigen (CBA). β-actin was used as a loading control. ( b ) Flow cytometry (FCM) analysis of IL-17 and interferon (IFN)-γ in STAT −/− naive T cells after in vitro differentiation with WT and Trif LPS2 MΦ in the presence of CBA. Representative results from five independent experiments are shown ( n =4 each, mean±s.e.m.). ( c ) Western blot analysis of transcription factors in WT naive T cells after co-culturing with WT MΦs and CBA in the presence of anti-IFNAR1 antibody. ( d ) FCM analysis of IL-17 and IFN-γ in STAT1 −/− naive T cells after co-culturing with Trif LPS2 MΦs in the presence of CBA and recombinant IL-27p28 (15 ng ml −1 ). Representative results from two independent experiments are shown ( n =4 each, mean±s.e.m.). ( e ) Western blot (top panel) and FCM (bottom panel) analysis of pSTAT1 in WT splenocytes after stimulation with IL-27p28 (15 ng ml −1 ) for the indicated period. For the FCM analysis, splenocytes were stained with CD3, CD4, and pSTAT1 after IL-27p28 stimulation. pSTAT1-positive cells in CD3 + CD4 + splenocytes are demonstrated. Results are representative of two independent experiments ( n =4 each, mean±s.e.m.). A full color version of this figure is available at the Mucosal Immunology journal online.
    Figure Legend Snippet: Interleukin (IL)-27p28-induced STAT1 activation regulates Th17-cell differentiation and plasticity. ( a ) Western blot analysis of transcription factors in wild-type (WT) naive T cells after co-culturing with MΦs in the presence of cecal bacterial antigen (CBA). β-actin was used as a loading control. ( b ) Flow cytometry (FCM) analysis of IL-17 and interferon (IFN)-γ in STAT −/− naive T cells after in vitro differentiation with WT and Trif LPS2 MΦ in the presence of CBA. Representative results from five independent experiments are shown ( n =4 each, mean±s.e.m.). ( c ) Western blot analysis of transcription factors in WT naive T cells after co-culturing with WT MΦs and CBA in the presence of anti-IFNAR1 antibody. ( d ) FCM analysis of IL-17 and IFN-γ in STAT1 −/− naive T cells after co-culturing with Trif LPS2 MΦs in the presence of CBA and recombinant IL-27p28 (15 ng ml −1 ). Representative results from two independent experiments are shown ( n =4 each, mean±s.e.m.). ( e ) Western blot (top panel) and FCM (bottom panel) analysis of pSTAT1 in WT splenocytes after stimulation with IL-27p28 (15 ng ml −1 ) for the indicated period. For the FCM analysis, splenocytes were stained with CD3, CD4, and pSTAT1 after IL-27p28 stimulation. pSTAT1-positive cells in CD3 + CD4 + splenocytes are demonstrated. Results are representative of two independent experiments ( n =4 each, mean±s.e.m.). A full color version of this figure is available at the Mucosal Immunology journal online.

    Techniques Used: Activation Assay, Cell Differentiation, Western Blot, Crocin Bleaching Assay, Flow Cytometry, Cytometry, In Vitro, Recombinant, Staining

    19) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    20) Product Images from "Interferon-? Induces Senescence in Normal Human Melanocytes"

    Article Title: Interferon-? Induces Senescence in Normal Human Melanocytes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0093232

    JAK2 and STAT1 activities are necessary for IFN-γ caused melanocyte senescence. Melanocytes were transfected with JAK1, JAK2, STAT1 siRNAs or scrambled control siRNA (Ctrl). After 48 h, cells were treated with or without 100 U/ml IFN-γ for additional 7 days. (A) Cell viability of melancytes was measured by MTS assay. (B) Protein level of p21 was evaluated by Western blot. β-actin was probed as the loading control. (C) Percentages of SA-β-gal-positive cells were determined based on microscopic analysis.
    Figure Legend Snippet: JAK2 and STAT1 activities are necessary for IFN-γ caused melanocyte senescence. Melanocytes were transfected with JAK1, JAK2, STAT1 siRNAs or scrambled control siRNA (Ctrl). After 48 h, cells were treated with or without 100 U/ml IFN-γ for additional 7 days. (A) Cell viability of melancytes was measured by MTS assay. (B) Protein level of p21 was evaluated by Western blot. β-actin was probed as the loading control. (C) Percentages of SA-β-gal-positive cells were determined based on microscopic analysis.

    Techniques Used: Transfection, MTS Assay, Western Blot

    21) Product Images from "IFN? mediates DUOX2 expression via a STAT-independent signaling pathway"

    Article Title: IFN? mediates DUOX2 expression via a STAT-independent signaling pathway

    Journal: Biochemical and biophysical research communications

    doi: 10.1016/j.bbrc.2010.04.004

    IFNγ induces JAK-mediated STAT1 phosphorylation in HBE1 cells
    Figure Legend Snippet: IFNγ induces JAK-mediated STAT1 phosphorylation in HBE1 cells

    Techniques Used:

    22) Product Images from "Prolonged TNFα primes fibroblast-like synoviocytes in a gene-specific manner by altering chromatin"

    Article Title: Prolonged TNFα primes fibroblast-like synoviocytes in a gene-specific manner by altering chromatin

    Journal: Arthritis & rheumatology (Hoboken, N.J.)

    doi: 10.1002/art.38871

    STAT1 amplifier function of chronic TNFα in RA FLS RA FLS were cultured for 3 days in the presence or absence of TNFα (10 ng/ml), which was added on the first day of culture and was not replenished ( A, D ). ( A ), On day 3 the cells were stimulated with IFNβ (1,000 U/ml) or IFNγ (100 U/ml) for 10-60 minutes and STAT1 tyrosine phosphorylation (pY) was measured by immunoblotting. The expression of STAT1 protein ( B ) and mRNA ( C ), upon TNFα time course stimulation, was measured by immunoblotting and qPCR. Values are the mean ±SEM and were normalized relative to mRNA for GAPDH. *= p
    Figure Legend Snippet: STAT1 amplifier function of chronic TNFα in RA FLS RA FLS were cultured for 3 days in the presence or absence of TNFα (10 ng/ml), which was added on the first day of culture and was not replenished ( A, D ). ( A ), On day 3 the cells were stimulated with IFNβ (1,000 U/ml) or IFNγ (100 U/ml) for 10-60 minutes and STAT1 tyrosine phosphorylation (pY) was measured by immunoblotting. The expression of STAT1 protein ( B ) and mRNA ( C ), upon TNFα time course stimulation, was measured by immunoblotting and qPCR. Values are the mean ±SEM and were normalized relative to mRNA for GAPDH. *= p

    Techniques Used: Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    23) Product Images from "Puerarin improves graft bone defect through microRNA-155-3p-mediated p53/TNF-α/STAT1 signaling pathway"

    Article Title: Puerarin improves graft bone defect through microRNA-155-3p-mediated p53/TNF-α/STAT1 signaling pathway

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2020.4595

    Puerarin regulated miR-155-3p-mediated p53/TNF-α/STAT1 signaling in BMSCs. (A) Expression of miR-155-3p in bone tissue in rats with bone graft defects. (B) IHC assay analyzed the expression of p53, TNF-α and STAT1 in bone tissue in the puerarin and PBS groups. (C) Expression of miR-155-3p in BMSCs. (D) Gene and (E) protein expression of p53, TNF-α and STAT1 in BMSCs. (F) Effects of miR-155 inhibitor on miR-155, p53, TNF-α and STAT1 mRNA expression in BMSCs. (G) Effect of miR-155 inhibitor on p53, TNF-α and STAT1 protein expression in BMSCs. Scale bars, 50 μ m. Data are expressed as the mean ± standard deviation. Each experiment was repeated at least three times. Student's t-test was used to evaluate the statistical significance of differences between two groups and one-way ANOVA followed by Tukey's test were performed for multiple groups. ** P
    Figure Legend Snippet: Puerarin regulated miR-155-3p-mediated p53/TNF-α/STAT1 signaling in BMSCs. (A) Expression of miR-155-3p in bone tissue in rats with bone graft defects. (B) IHC assay analyzed the expression of p53, TNF-α and STAT1 in bone tissue in the puerarin and PBS groups. (C) Expression of miR-155-3p in BMSCs. (D) Gene and (E) protein expression of p53, TNF-α and STAT1 in BMSCs. (F) Effects of miR-155 inhibitor on miR-155, p53, TNF-α and STAT1 mRNA expression in BMSCs. (G) Effect of miR-155 inhibitor on p53, TNF-α and STAT1 protein expression in BMSCs. Scale bars, 50 μ m. Data are expressed as the mean ± standard deviation. Each experiment was repeated at least three times. Student's t-test was used to evaluate the statistical significance of differences between two groups and one-way ANOVA followed by Tukey's test were performed for multiple groups. ** P

    Techniques Used: Expressing, Immunohistochemistry, Standard Deviation

    24) Product Images from "Anti-Allergic and Anti-Inflammatory Effects of Kuwanon G and Morusin on MC/9 Mast Cells and HaCaT Keratinocytes"

    Article Title: Anti-Allergic and Anti-Inflammatory Effects of Kuwanon G and Morusin on MC/9 Mast Cells and HaCaT Keratinocytes

    Journal: Molecules

    doi: 10.3390/molecules24020265

    Effect of morusin on TNF-α and IFN-γ-induced STAT1 ( a ) and NF-κB p65 ( b ) phosphorylation in HaCaT keratinocytes. Expression of total and phosphorylated STAT1 (pSTAT1) and NF-κB p65 (pNF-κB p65) was determined by Western blotting. Silymarin was used as the positive control. Data are expressed as the mean ± SEM (n = 2). # p
    Figure Legend Snippet: Effect of morusin on TNF-α and IFN-γ-induced STAT1 ( a ) and NF-κB p65 ( b ) phosphorylation in HaCaT keratinocytes. Expression of total and phosphorylated STAT1 (pSTAT1) and NF-κB p65 (pNF-κB p65) was determined by Western blotting. Silymarin was used as the positive control. Data are expressed as the mean ± SEM (n = 2). # p

    Techniques Used: Expressing, Western Blot, Positive Control

    Effect of kuwanon G on TNF-α and IFN-γ-induced STAT1 ( a ) and NF-κB p65 ( b ) phosphorylation in HaCaT keratinocytes. Expression of total and phosphorylated STAT1 (pSTAT1) and NF-κB p65 (pNF-κB p65) was determined by Western blotting. Silymarin was used as the positive control. Data are expressed as the mean ± SEM ( n = 2). # p
    Figure Legend Snippet: Effect of kuwanon G on TNF-α and IFN-γ-induced STAT1 ( a ) and NF-κB p65 ( b ) phosphorylation in HaCaT keratinocytes. Expression of total and phosphorylated STAT1 (pSTAT1) and NF-κB p65 (pNF-κB p65) was determined by Western blotting. Silymarin was used as the positive control. Data are expressed as the mean ± SEM ( n = 2). # p

    Techniques Used: Expressing, Western Blot, Positive Control

    25) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    26) Product Images from "THE ROLE OF CYTOKINES IN UBD PROMOTER REGULATION AND MALLORY-DENK BODY-LIKE AGGRESOMES"

    Article Title: THE ROLE OF CYTOKINES IN UBD PROMOTER REGULATION AND MALLORY-DENK BODY-LIKE AGGRESOMES

    Journal: Experimental and molecular pathology

    doi: 10.1016/j.yexmp.2010.04.001

    Analysis of the phosphorylation levels of ERK/p42/44, STAT1 and STAT3. TNFa was able to induce the phosphorylation of p42/44. IFNg increased the phosphorylation of p42/44, STAT1 and STAT3. The combination of the two cytokines (I+T) had a synergistic effect
    Figure Legend Snippet: Analysis of the phosphorylation levels of ERK/p42/44, STAT1 and STAT3. TNFa was able to induce the phosphorylation of p42/44. IFNg increased the phosphorylation of p42/44, STAT1 and STAT3. The combination of the two cytokines (I+T) had a synergistic effect

    Techniques Used:

    27) Product Images from "The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney"

    Article Title: The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1005734

    Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P
    Figure Legend Snippet: Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P

    Techniques Used: Expressing, Activation Assay, Staining

    Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (
    Figure Legend Snippet: Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (

    Techniques Used: Activity Assay, Expressing

    Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).
    Figure Legend Snippet: Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).

    Techniques Used: Activation Assay, Expressing, Microarray

    28) Product Images from "Transgenic Expression of Interferon-? in Mouse Stomach Leads to Inflammation, Metaplasia, and Dysplasia"

    Article Title: Transgenic Expression of Interferon-? in Mouse Stomach Leads to Inflammation, Metaplasia, and Dysplasia

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2012.08.017

    Activation of STAT1 and STAT3 in the corpus of H/K-IFN-γ mice. A–C: Immunostaining for STAT1 reveals marked upregulation in H/K-IFN-γ mouse with negligible expression in control stomach. Higher magnification in C shows a subset
    Figure Legend Snippet: Activation of STAT1 and STAT3 in the corpus of H/K-IFN-γ mice. A–C: Immunostaining for STAT1 reveals marked upregulation in H/K-IFN-γ mouse with negligible expression in control stomach. Higher magnification in C shows a subset

    Techniques Used: Activation Assay, Mouse Assay, Immunostaining, Expressing

    29) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    30) Product Images from "Prolonged TNFα primes fibroblast-like synoviocytes in a gene-specific manner by altering chromatin"

    Article Title: Prolonged TNFα primes fibroblast-like synoviocytes in a gene-specific manner by altering chromatin

    Journal: Arthritis & rheumatology (Hoboken, N.J.)

    doi: 10.1002/art.38871

    STAT1 amplifier function of chronic TNFα in RA FLS RA FLS were cultured for 3 days in the presence or absence of TNFα (10 ng/ml), which was added on the first day of culture and was not replenished ( A, D ). ( A ), On day 3 the cells were stimulated with IFNβ (1,000 U/ml) or IFNγ (100 U/ml) for 10-60 minutes and STAT1 tyrosine phosphorylation (pY) was measured by immunoblotting. The expression of STAT1 protein ( B ) and mRNA ( C ), upon TNFα time course stimulation, was measured by immunoblotting and qPCR. Values are the mean ±SEM and were normalized relative to mRNA for GAPDH. *= p
    Figure Legend Snippet: STAT1 amplifier function of chronic TNFα in RA FLS RA FLS were cultured for 3 days in the presence or absence of TNFα (10 ng/ml), which was added on the first day of culture and was not replenished ( A, D ). ( A ), On day 3 the cells were stimulated with IFNβ (1,000 U/ml) or IFNγ (100 U/ml) for 10-60 minutes and STAT1 tyrosine phosphorylation (pY) was measured by immunoblotting. The expression of STAT1 protein ( B ) and mRNA ( C ), upon TNFα time course stimulation, was measured by immunoblotting and qPCR. Values are the mean ±SEM and were normalized relative to mRNA for GAPDH. *= p

    Techniques Used: Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    31) Product Images from "Proteoform-Specific Insights into Cellular Proteome Regulation *"

    Article Title: Proteoform-Specific Insights into Cellular Proteome Regulation *

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1074/mcp.O116.058438

    Analysis of STAT1 proteoforms induced by hRSV infection of A549 cells. Panel A presents the observed focusing profile for the STAT1 protein group. Focusing for the five uninfected lysates are represented in blue and focusing for the five hRSV-infected lysates are represented in red. Arrows represent the predicted focusing for all proteoform sequences belonging to the STAT1 protein group. Panels B and C present data obtained from 1D-Western blots of four independent biological replicate sets of uninfected (Mock) and hRSV-infected (hRSV) A549 cell lysates. Panel B , is a representative image of a 1D-Western blot obtained with equal quantities of protein from Mock and hRSV lysates from one replicate. Stat1-α and –β are evident as green bands with mobilities corresponding to 91 and 84 kDa, respectively. The red band represents β-actin used as a loading control. Quantitative data are presented in panel C for data obtained from the complete sample set with the Stat1-α and –β proteoforms presented on the left and right, respectively. Integrated intensities are presented as the mean ± S.E. ( n = 4). Statistical analysis was performed using a paired, two-tailed Student's t test. p values are presented in both panels. Western blots after 2D-separation of equal quantities of protein from one replicate of D Mock and E hRSV lysates. Arrows pointing to the right indicate different proteoforms of Stat1-α and to the left indicate different proteoforms of Stat1-β.
    Figure Legend Snippet: Analysis of STAT1 proteoforms induced by hRSV infection of A549 cells. Panel A presents the observed focusing profile for the STAT1 protein group. Focusing for the five uninfected lysates are represented in blue and focusing for the five hRSV-infected lysates are represented in red. Arrows represent the predicted focusing for all proteoform sequences belonging to the STAT1 protein group. Panels B and C present data obtained from 1D-Western blots of four independent biological replicate sets of uninfected (Mock) and hRSV-infected (hRSV) A549 cell lysates. Panel B , is a representative image of a 1D-Western blot obtained with equal quantities of protein from Mock and hRSV lysates from one replicate. Stat1-α and –β are evident as green bands with mobilities corresponding to 91 and 84 kDa, respectively. The red band represents β-actin used as a loading control. Quantitative data are presented in panel C for data obtained from the complete sample set with the Stat1-α and –β proteoforms presented on the left and right, respectively. Integrated intensities are presented as the mean ± S.E. ( n = 4). Statistical analysis was performed using a paired, two-tailed Student's t test. p values are presented in both panels. Western blots after 2D-separation of equal quantities of protein from one replicate of D Mock and E hRSV lysates. Arrows pointing to the right indicate different proteoforms of Stat1-α and to the left indicate different proteoforms of Stat1-β.

    Techniques Used: Infection, Western Blot, Two Tailed Test

    32) Product Images from "Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer"

    Article Title: Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer

    Journal: Oncoimmunology

    doi: 10.1080/2162402X.2017.1288330

    Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of STAT1, IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.
    Figure Legend Snippet: Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of STAT1, IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.

    Techniques Used: Expressing, Activation Assay, CTL Assay

    33) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    34) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    35) Product Images from "Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia"

    Article Title: Role of CTLA4 in the Proliferation and Survival of Chronic Lymphocytic Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070352

    Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).
    Figure Legend Snippet: Differential expression of CTLA4 and associated molecules in primary CLL cells. Panel I: Differential expression of CTLA4 and associated molecules in high-CD38/low-CTLA4 and low-CD38/high-CTLA4 CLL subgroups as determined by microarray data. Subpanel A: Mean normalized expression levels of CTLA4 in the CD38-low group compared to the CD38-high group. Subpanels B–D: Mean normalized expression levels of STAT1, NFATC2, and c-Fos (respectively) in the CTLA4-high group compared to CTLA4-low group. Panel II: Overexpression of downstream molecules in BCR proliferation signaling in two prognostic CLL subgroups, as determined by real-time PCR. Subpanels A–C: Real-time PCR showing gene expression for STAT1, NFATC2, and c-Fos (fold change) in the high-CTLA4 CLL group compared to the low-CTLA4 CLL group, respectively. Subpanels D–F: Real-time PCR showing gene expression of STAT1, NFATC2, and c-Fos (fold change) in chromosomal abnormality subgroups, respectively (good outcome includes 13q14 deletion and normal karyotype, while poor outcome includes trisomy12, 11q deletion, and 17p deletion chromosomal abnormality).

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction

    Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.
    Figure Legend Snippet: Upregulation of STAT1/phospho-STAT1, NFATC2, c-Fos/phospho-c-Fos, and c-Myc in CTLA4-downregulated CLL (low CD38/high CTLA4) cells as determined by RT-PCR, real-time PCR, and/or Western blotting. Panels A: Semi-quantitative RT-PCR showing that downregulation of CTLA4 by AS in CLL cells leads to up-regulation of STAT1, NFATC2, c-Fos, and c-Myc in CLL. Panel B: Real-time PCR results from four patient samples showing a significantly higher expression of c-Myc in CTLA4-downregulated CLL cells compared to control CLL cells or CLL cells treated with irrelevant AS. Panel C–D: Western blot results showing up-regulation and quantification of the expression of STAT1 and its phosphorylation, c-Fos and its phosphorylation, NFATC2, and c-Myc at the protein level in CTLA4 down-regulated CLL patient samples (n = 3). *p indicates the statistical difference between control CLL cells and CTLA4-downregulated CLL cells. β-Actin was used as a control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Expressing

    Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.
    Figure Legend Snippet: Hypothetical model for the role of CTLA4 in CLL cell proliferation/survival. Panel A: When CLL cells express low CD38, but high CTLA4, CTLA4 inhibits the CD38/BCR signaling pathway at multiple levels. CTLA4 downregulates NFATC2 and proliferation-associated molecules such as c-Fos and c-Myc. Downregulation of NFATC2 may also be associated with an autoregulatory loop for CTLA4, which would downregulate CTLA4 transcription. CTLA4 also downregulates the expression of Bcl-2, thus decreasing the survival of CLL cells. CTLA4 inhibits the expression of STAT1, thus deregulating the JAK/STAT pathway and inhibiting CLL cell growth. Panel B: When CLL cells express high CD38, but low CTLA4, activated CD38/BCR signaling upregulates downstream molecules in the pathway, such as NFATC2, c-Fos, and Bcl-2. These molecules will increase proliferation and survival of CLL cells. Low expression of CTLA4 does not interfere with the expression of STAT1, which favors CLL cell growth.

    Techniques Used: Expressing

    36) Product Images from "The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney"

    Article Title: The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1005734

    Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P
    Figure Legend Snippet: Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P

    Techniques Used: Expressing, Activation Assay, Staining

    Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (
    Figure Legend Snippet: Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (

    Techniques Used: Activity Assay, Expressing

    Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).
    Figure Legend Snippet: Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).

    Techniques Used: Activation Assay, Expressing, Microarray

    37) Product Images from "The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney"

    Article Title: The Inflammatory Transcription Factors NFκB, STAT1 and STAT3 Drive Age-Associated Transcriptional Changes in the Human Kidney

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1005734

    Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P
    Figure Legend Snippet: Correlations of the activities of STAT1, STAT3, NFκB and macrophage abundance in individual kidneys. A. In this heat map the rows indicate estimated macrophage abundance based on macrophage-specific transcript expression, or level of activation of each transcription factor, based on the averaged expression of transcription factor direct target genes (see Methods ). The colors indicate relative levels of macrophage abundance or transcription factor target gene expression compared to other individuals of that age; red columns represent individuals with higher expression of macrophage-specific transcripts or transcription factor target genes for their age (older than expected at the gene expression level), and blue columns represent individuals with lower expression levels of macrophage markers of transcription factor target genes for their age (more youthful at the gene expression level). The columns are clustered such that individuals with values that are high or low for their age appear together. B. Macrophage infiltration in the kidney increases with age: images of renal cortex samples showing CD163 staining (green) with DAPI counterstaining (blue). Shown is a representative example of a young renal cortex (left) and an old renal cortex (right). The boxplot quantifies macrophage infiltration. The relative abundance of macrophages was defined as the fraction of CD163 + cells/all DAPI + cells The boxes indicate 25th and 75th percentiles for the group, and the lines indicate maximum, median and minimum values. ** P

    Techniques Used: Expressing, Activation Assay, Staining

    Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (
    Figure Legend Snippet: Increased activity of STAT1, STAT3 and NFκB transcription factors during kidney aging. A. Boxplots showing significantly increased mRNA expression of STAT1 and STAT3 in old ( > 65 years, n = 42) versus young (

    Techniques Used: Activity Assay, Expressing

    Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).
    Figure Legend Snippet: Activation of STAT1, STAT3 or NFκB by inflammatory cytokines recapitulates kidney aging-related gene expression patterns in human renal epithelial cells. A. The left column of the heat map shows the log 2 fold-changes of 40 direct targets of STAT1 following IFNγ stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these STAT1 direct targets during kidney aging. B. The left column of heat map shows the log 2 fold-changes of 43 direct targets of STAT3 following IL-6 stimulation of HK-2 cells from microarray expression profiling experiments. The middle column shows changes in expression following treatment with the STAT3 inhibitor S3I-201. The right column shows the corresponding log 2 -adjusted beta coefficient (age-slope) during kidney aging. Aging gene expression data are from [ 1 ]. C. The left column of the heat map shows the log 2 fold-changes of 43 direct targets of NFκB following TNFα stimulation of HK-2 cells from microarray expression profiling experiments. The right column of the heat map shows the corresponding log 2 -adjusted beta coefficient (age-slope) for these NFκB direct targets during kidney aging. Yellow indicates increased gene expression (positive fold-change or age-slope) and blue indicates decreased gene expression (negative-fold change or age-slope).

    Techniques Used: Activation Assay, Expressing, Microarray

    38) Product Images from "Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression"

    Article Title: Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095627

    rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p
    Figure Legend Snippet: rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p

    Techniques Used: Immunofluorescence, Western Blot, Flow Cytometry, MANN-WHITNEY

    Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.
    Figure Legend Snippet: Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.

    Techniques Used:

    39) Product Images from "Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression"

    Article Title: Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095627

    rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p
    Figure Legend Snippet: rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p

    Techniques Used: Immunofluorescence, Western Blot, Flow Cytometry, MANN-WHITNEY

    Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.
    Figure Legend Snippet: Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.

    Techniques Used:

    40) Product Images from "Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression"

    Article Title: Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095627

    rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p
    Figure Legend Snippet: rCore alters JAK-STAT proteins. A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p

    Techniques Used: Immunofluorescence, Western Blot, Flow Cytometry, MANN-WHITNEY

    Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.
    Figure Legend Snippet: Paradigm Model of HCV Core acting on pDCs. A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.

    Techniques Used:

    Related Articles

    Transduction:

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. Overexpression of STAT3 reversed the co-treatment of STAT3 shRNA and GEN on M2 phenotype of THP-1 macrophages induced by co-culture of THP-1 macrophages expressing Ad-shSTAT3 were transduced with Ad-STAT3 and co-cultured with SKOV3-derived OCSLCs. .. Ad-shSTAT3: The cells transduced with adenovirus expressing shSTAT3.

    Immunohistochemistry:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    shRNA:

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. Overexpression of STAT3 reversed the co-treatment of STAT3 shRNA and GEN on M2 phenotype of THP-1 macrophages induced by co-culture of THP-1 macrophages expressing Ad-shSTAT3 were transduced with Ad-STAT3 and co-cultured with SKOV3-derived OCSLCs. .. Ad-shSTAT3: The cells transduced with adenovirus expressing shSTAT3.

    Inverted Microscopy:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Stable Transfection:

    Article Title: Lentivirus-mediated siRNA knockdown of SPHK1 inhibits proliferation and tumorigenesis of neuroblastoma
    Article Snippet: .. With the aim to identify the role of STAT3 in suppression of NB progression via SPHK1 down regulation, we first transfected STAT3 expression vector (pcDNA3.1 (+)-STAT3) into SH-SY5Y cells (stably expressing SPHK1 siRNA), which helped us evaluate the STAT3 expression level in these cells to almost the same level of blank control (SPHK1 siRNA untreated cells) ( ). .. Interestingly, we found that ectopic overexpression of STAT3 could reverse the inhibition effect caused by SPHK1 siRNA expression in SH-SY5Y cells.

    CCK-8 Assay:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Flow Cytometry:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Co-Culture Assay:

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. Overexpression of STAT3 reversed the co-treatment of STAT3 shRNA and GEN on M2 phenotype of THP-1 macrophages induced by co-culture of THP-1 macrophages expressing Ad-shSTAT3 were transduced with Ad-STAT3 and co-cultured with SKOV3-derived OCSLCs. .. Ad-shSTAT3: The cells transduced with adenovirus expressing shSTAT3.

    Cytometry:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Lipopolysaccharide shock reveals the immune function of indoleamine 2,3-dioxygenase 2 through the regulation of IL-6/stat3 signalling
    Article Snippet: .. The phosphorylated stat3 levels and stat3 protein levels were examined using a stat3 (pY705) and total stat3 ELISA Kit (ab126459) (Abcam, Cambridge, UK). .. A sandwich ELISA system is an in vitro enzyme-linked immunosorbent assay for the measurement of phospho-stat3 (Tyr705) and total stat3.

    Activation Assay:

    Article Title: EGFRvIII-Stat5 Signaling Enhances Glioblastoma Cell Migration and Survival
    Article Snippet: .. Since we observed higher Stat5 activation in the GBM rim cells, we next investigated the role of Stat5 in the regulation of GBM migration. .. We tested three different siRNAs targeting each of the Stat isoforms and chose the siRNAs displaying the highest specific mRNA depletion for functional studies ( ).

    Recombinant:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Transfection:

    Article Title: Lentivirus-mediated siRNA knockdown of SPHK1 inhibits proliferation and tumorigenesis of neuroblastoma
    Article Snippet: .. With the aim to identify the role of STAT3 in suppression of NB progression via SPHK1 down regulation, we first transfected STAT3 expression vector (pcDNA3.1 (+)-STAT3) into SH-SY5Y cells (stably expressing SPHK1 siRNA), which helped us evaluate the STAT3 expression level in these cells to almost the same level of blank control (SPHK1 siRNA untreated cells) ( ). .. Interestingly, we found that ectopic overexpression of STAT3 could reverse the inhibition effect caused by SPHK1 siRNA expression in SH-SY5Y cells.

    Migration:

    Article Title: EGFRvIII-Stat5 Signaling Enhances Glioblastoma Cell Migration and Survival
    Article Snippet: .. Since we observed higher Stat5 activation in the GBM rim cells, we next investigated the role of Stat5 in the regulation of GBM migration. .. We tested three different siRNAs targeting each of the Stat isoforms and chose the siRNAs displaying the highest specific mRNA depletion for functional studies ( ).

    Expressing:

    Article Title: Lentivirus-mediated siRNA knockdown of SPHK1 inhibits proliferation and tumorigenesis of neuroblastoma
    Article Snippet: .. With the aim to identify the role of STAT3 in suppression of NB progression via SPHK1 down regulation, we first transfected STAT3 expression vector (pcDNA3.1 (+)-STAT3) into SH-SY5Y cells (stably expressing SPHK1 siRNA), which helped us evaluate the STAT3 expression level in these cells to almost the same level of blank control (SPHK1 siRNA untreated cells) ( ). .. Interestingly, we found that ectopic overexpression of STAT3 could reverse the inhibition effect caused by SPHK1 siRNA expression in SH-SY5Y cells.

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. As indicated in Fig. a, overexpression of STAT3 abrogated the depletion of IL-8 combined with GEN decreased the expression levels of CD44 and CD133 in SKOV3 cells induced by Co-CM. .. Figure b and c showed that overexpression of STAT3 reduced the depletion of IL-8 combined with GEN inhibited the self-renewal ability and in vitro tumorigenic capabilities in SKOV3 cells induced by Co-CM.

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. Overexpression of STAT3 reversed the co-treatment of STAT3 shRNA and GEN on M2 phenotype of THP-1 macrophages induced by co-culture of THP-1 macrophages expressing Ad-shSTAT3 were transduced with Ad-STAT3 and co-cultured with SKOV3-derived OCSLCs. .. Ad-shSTAT3: The cells transduced with adenovirus expressing shSTAT3.

    Staining:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Western Blot:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Over Expression:

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. As indicated in Fig. a, overexpression of STAT3 abrogated the depletion of IL-8 combined with GEN decreased the expression levels of CD44 and CD133 in SKOV3 cells induced by Co-CM. .. Figure b and c showed that overexpression of STAT3 reduced the depletion of IL-8 combined with GEN inhibited the self-renewal ability and in vitro tumorigenic capabilities in SKOV3 cells induced by Co-CM.

    Article Title: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis
    Article Snippet: .. Overexpression of STAT3 reversed the co-treatment of STAT3 shRNA and GEN on M2 phenotype of THP-1 macrophages induced by co-culture of THP-1 macrophages expressing Ad-shSTAT3 were transduced with Ad-STAT3 and co-cultured with SKOV3-derived OCSLCs. .. Ad-shSTAT3: The cells transduced with adenovirus expressing shSTAT3.

    other:

    Article Title: EGFRvIII-Stat5 Signaling Enhances Glioblastoma Cell Migration and Survival
    Article Snippet: We next assessed the clinical relevance of differential intratumoral localization of the Stat transcription factors by measuring levels of activated Stat3 and Stat5 on a GBM invasion TMA( ).

    Plasmid Preparation:

    Article Title: Establishment of malignantly transformed dendritic cell line SU3-ihDCTC induced by Glioma stem cells and study on its sensitivity to resveratrol
    Article Snippet: .. The RFP lentiviral vector was purchased from Shanghai Innovation Biotechnology Co., Ltd.; hamster anti-mouse CD11c antibody from eBioscience Corporation, US; APC-labeled anti-mouse CD11c antibody and APC-labeled anti-mouse CD80 antibody from Biolegend Corporation, US; recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin 4 (rmIL-4) from Peprotech Corporation; rabbit anti-mouse α signal protein (SIRP-α) antibody from Abcam, Inc.; CCK-8 reagent from Dojindo Chemical Technology Co., Ltd.; immunohistochemical staining and Western Blot primary antibody reagents: antibodies against IL-6, STAT3, p-STAT3, NF-κB and p-NF-κB were purchased from Abcam, Inc. DMEM medium and fetal bovine serum were purchased from Hyclone Laboratories, Inc., US; flow cytometer from Beckman Coulter, US; fluorescent inverted microscope from Olympus Corporation, Japan; microplate reader from Tecan, Switzerland. .. Freezing microtome was purchased from LEICA, Germany and cell incubator from SANYO, Japan.

    Article Title: Lentivirus-mediated siRNA knockdown of SPHK1 inhibits proliferation and tumorigenesis of neuroblastoma
    Article Snippet: .. With the aim to identify the role of STAT3 in suppression of NB progression via SPHK1 down regulation, we first transfected STAT3 expression vector (pcDNA3.1 (+)-STAT3) into SH-SY5Y cells (stably expressing SPHK1 siRNA), which helped us evaluate the STAT3 expression level in these cells to almost the same level of blank control (SPHK1 siRNA untreated cells) ( ). .. Interestingly, we found that ectopic overexpression of STAT3 could reverse the inhibition effect caused by SPHK1 siRNA expression in SH-SY5Y cells.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Abcam p stat1
    The protein levels of <t>p-STAT1;</t> p-STAT3; p-STAT5; and STAT-1, STAT-3, and STAT5 in normal, vehicle, and 0.5% MMB groups, respectively. ( A ) Western blot and ( B ) quantitative analysis of dot intensity. Actin was used as a reference protein. Values are mean ± SD ( n = 3). * * p
    P Stat1, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p stat1/product/Abcam
    Average 92 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    p stat1 - by Bioz Stars, 2020-11
    92/100 stars
      Buy from Supplier

    stat1  (Abcam)
    99
    Abcam stat1
    Inhibitory effect of SARS-CoV PLPro on TLR7 agonist-induced activation of type I IFN signaling. ISRE-driven luciferase reporter activity and the mRNA levels of PKR and IRF7 were determined 4 h post-IMQ treatment. ISRE-driven firefly luciferase activity was normalized by Renilla luciferase activity ( A ). Relative mRNA levels of PKR ( B ) and IRF7 ( C ) were normalized by GAPDH mRNA, presented as a relative ratio. In addition, the activated status of <t>STAT1</t> was examined using Western blot with anti-phospho-STAT1 (Tyr701) antibodies ( D ). * p -Value
    Stat1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stat1/product/Abcam
    Average 99 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    stat1 - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    96
    Abcam ph stat1
    Kaplan-Meier survival curves (Log rank) of cancer specific survival A. Total <t>STAT1</t> tumour cell expression and B. Ph-STAT1 tumour cell expression. C. Total STAT3 tumour cell expression and D. Ph-STAT3 tumour cell expression.
    Ph Stat1, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ph stat1/product/Abcam
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ph stat1 - by Bioz Stars, 2020-11
    96/100 stars
      Buy from Supplier

    Image Search Results


    The protein levels of p-STAT1; p-STAT3; p-STAT5; and STAT-1, STAT-3, and STAT5 in normal, vehicle, and 0.5% MMB groups, respectively. ( A ) Western blot and ( B ) quantitative analysis of dot intensity. Actin was used as a reference protein. Values are mean ± SD ( n = 3). * * p

    Journal: International Journal of Molecular Sciences

    Article Title: Topical Application of JAK1/JAK2 Inhibitor Momelotinib Exhibits Significant Anti-Inflammatory Responses in DNCB-Induced Atopic Dermatitis Model Mice

    doi: 10.3390/ijms19123973

    Figure Lengend Snippet: The protein levels of p-STAT1; p-STAT3; p-STAT5; and STAT-1, STAT-3, and STAT5 in normal, vehicle, and 0.5% MMB groups, respectively. ( A ) Western blot and ( B ) quantitative analysis of dot intensity. Actin was used as a reference protein. Values are mean ± SD ( n = 3). * * p

    Article Snippet: Stat1; p-Stat1; Stat3; p-Stat3; Stat5; p-Stat5 were provided by Abcom (Cambridge, UK).

    Techniques: Western Blot

    Inhibitory effect of SARS-CoV PLPro on TLR7 agonist-induced activation of type I IFN signaling. ISRE-driven luciferase reporter activity and the mRNA levels of PKR and IRF7 were determined 4 h post-IMQ treatment. ISRE-driven firefly luciferase activity was normalized by Renilla luciferase activity ( A ). Relative mRNA levels of PKR ( B ) and IRF7 ( C ) were normalized by GAPDH mRNA, presented as a relative ratio. In addition, the activated status of STAT1 was examined using Western blot with anti-phospho-STAT1 (Tyr701) antibodies ( D ). * p -Value

    Journal: International Journal of Molecular Sciences

    Article Title: SARS Coronavirus Papain-Like Protease Inhibits the TLR7 Signaling Pathway through Removing Lys63-Linked Polyubiquitination of TRAF3 and TRAF6

    doi: 10.3390/ijms17050678

    Figure Lengend Snippet: Inhibitory effect of SARS-CoV PLPro on TLR7 agonist-induced activation of type I IFN signaling. ISRE-driven luciferase reporter activity and the mRNA levels of PKR and IRF7 were determined 4 h post-IMQ treatment. ISRE-driven firefly luciferase activity was normalized by Renilla luciferase activity ( A ). Relative mRNA levels of PKR ( B ) and IRF7 ( C ) were normalized by GAPDH mRNA, presented as a relative ratio. In addition, the activated status of STAT1 was examined using Western blot with anti-phospho-STAT1 (Tyr701) antibodies ( D ). * p -Value

    Article Snippet: Lysate from IMQ-treated cells was used for the Western blotting assay; the resulting blots were probed with specific primary antibodies against IRF3, phosphor-IRF3, STAT1, phospho-STAT1(Tyr701), NF-κB p65, phospho-NF-κB p65, p38 MAPK, phospho-p38 MAPK, c-Jun, phospho-c-Jun, TBK1, phospho-TBK1, TRAF3, TRAF6 and anti-β-actin mAb (Abcam, Cambridge, UK).

    Techniques: Activation Assay, Luciferase, Activity Assay, Western Blot

    Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of STAT1, IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.

    Journal: Oncoimmunology

    Article Title: Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer

    doi: 10.1080/2162402X.2017.1288330

    Figure Lengend Snippet: Working model. (A) The immune-activated phenotype of CRC is characterized by low expression of miR-34a and miR-93, activation of interferon signaling and expression of STAT1, IRF-1 and IRF-5 in the tumor microenvironment. Tumors of this phenotype are strongly infiltrated by CTLs releasing cytotoxic effector molecules, frequently have a microsatellite instable genotype and show upregulation of ICAM-1 on tumor cells. Patients with a high ISG-score have a significantly reduced risk of presenting with a distant metastasis. (B) The immune-quiescent phenotype of CRC is characterized by silencing of ISG through miR-34a and miR-93 in the tumor microenvironment and reduced of T-cell activation. Expression of cytotoxic effector molecules in CTL is infrequent. Lack of ICAM-1 may make tumors of this phenotype less amenable to CTL-infiltration and resistant to immune-mediated tumor destruction. Tumors of this phenotype frequently have a microsatellite stable genotype. Patients with a low ISG-score have a significantly increased risk of presenting with distant metastasis.

    Article Snippet: Cutting edge: differentiation of antitumor CTL in vivo requires host expression of Stat1 .

    Techniques: Expressing, Activation Assay, CTL Assay

    Kaplan-Meier survival curves (Log rank) of cancer specific survival A. Total STAT1 tumour cell expression and B. Ph-STAT1 tumour cell expression. C. Total STAT3 tumour cell expression and D. Ph-STAT3 tumour cell expression.

    Journal: Oncotarget

    Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

    doi: 10.18632/oncotarget.12730

    Figure Lengend Snippet: Kaplan-Meier survival curves (Log rank) of cancer specific survival A. Total STAT1 tumour cell expression and B. Ph-STAT1 tumour cell expression. C. Total STAT3 tumour cell expression and D. Ph-STAT3 tumour cell expression.

    Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

    Techniques: Expressing

    Sections of invasive ductal beast carcinomas showing IHC expression levels of ph-STAT1 (first row) and ph-STAT3 (second row) No appreciable expression was detected in the negative controls of ph-STAT1 A. and ph-STAT3 B . C-H. show the staining intensity of the STAT1 and STAT3 expression as low (C and D), moderate (E and F), and strong (G and H). Original magnification, 20×. Scale bars = 100 μm (A-F), 10 μm (G and H).

    Journal: Oncotarget

    Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

    doi: 10.18632/oncotarget.12730

    Figure Lengend Snippet: Sections of invasive ductal beast carcinomas showing IHC expression levels of ph-STAT1 (first row) and ph-STAT3 (second row) No appreciable expression was detected in the negative controls of ph-STAT1 A. and ph-STAT3 B . C-H. show the staining intensity of the STAT1 and STAT3 expression as low (C and D), moderate (E and F), and strong (G and H). Original magnification, 20×. Scale bars = 100 μm (A-F), 10 μm (G and H).

    Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

    Techniques: Immunohistochemistry, Expressing, Staining

    Kaplan-Meier survival curves (Log rank) of ph-STAT1 in different molecular subtypes Only Luminal A (n=174, 45%) shows significant association between high tumour cell expression of ph-STAT1 (n=121, 32%) and improved cancer specific survival

    Journal: Oncotarget

    Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

    doi: 10.18632/oncotarget.12730

    Figure Lengend Snippet: Kaplan-Meier survival curves (Log rank) of ph-STAT1 in different molecular subtypes Only Luminal A (n=174, 45%) shows significant association between high tumour cell expression of ph-STAT1 (n=121, 32%) and improved cancer specific survival

    Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

    Techniques: Expressing