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    Name:
    Animal Free Recombinant Human IL 17F
    Description:
    Animal Free Recombinant Human IL 17F Syn Interleukin 17F Source E coli Formulation Lyophilized
    Catalog Number:
    AF-200-25-100UG
    Price:
    660.00
    Category:
    Recombinant Proteins
    Source:
    E.coli
    Purity:
    98.0
    Quantity:
    100ug
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    Structured Review

    PeproTech il 17f
    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) <t>IL-17F</t> (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Animal Free Recombinant Human IL 17F Syn Interleukin 17F Source E coli Formulation Lyophilized
    https://www.bioz.com/result/il 17f/product/PeproTech
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 17f - by Bioz Stars, 2021-05
    92/100 stars

    Images

    1) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    2) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    3) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    4) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    5) Product Images from "IL-17F Promotes Tissue Injury in Autoimmune Kidney Diseases"

    Article Title: IL-17F Promotes Tissue Injury in Autoimmune Kidney Diseases

    Journal: Journal of the American Society of Nephrology : JASN

    doi: 10.1681/ASN.2015101077

    IL-17F promotes tissue injury by the induction of neutrophil-attracting chemokines. Diagram depicting the present hypothesis of the chain of events leading to the recruitment of tissue-destructive neutrophils by IL-17F. CD4 + T cell- and γδ T cell-derived IL-17F induces the expression and production of chemokines CXCL1 and CXCL5, which then attract CXCR2-expressing neutrophils to the inflamed kidney, leading to the destruction of regular kidney tissue and consequent loss of renal function.
    Figure Legend Snippet: IL-17F promotes tissue injury by the induction of neutrophil-attracting chemokines. Diagram depicting the present hypothesis of the chain of events leading to the recruitment of tissue-destructive neutrophils by IL-17F. CD4 + T cell- and γδ T cell-derived IL-17F induces the expression and production of chemokines CXCL1 and CXCL5, which then attract CXCR2-expressing neutrophils to the inflamed kidney, leading to the destruction of regular kidney tissue and consequent loss of renal function.

    Techniques Used: Derivative Assay, Expressing

    IL-17F drives renal tissue injury by mediating neutrophil infiltration. (A) Representative photographs of PAS-stained kidney sections from control, nephritic wild-type, and nephritic IL-17F–deficient mice at day 8 of NTN (original magnification ×400). (B) Quantification of glomerular crescent formation, interstitial area, and glomerular sclerosis of controls ( n =6), nephritic wild-type ( n =11), and nephritic IL-17F–deficient mice ( n =11) 8 days after induction of nephritis. (C) BUN levels, serum creatinine, and ACR of the aforementioned groups 8 days after induction of nephritis. (D) Quantification of tubulointerstitial GR-1 + cells, F4/80 + cells, and glomerular MAC-2 + cells in the aforementioned groups 8 days after induction of nephritis. (E) Representative FACS plots showing renal neutrophils (defined as CD45 + CD11b + Ly6G + cells) in nephritic wild-type and nephritic IL-17F–deficient mice and (F) quantification of FACS analysis of renal and splenic neutrophils of wild-type controls ( n =3–4), IL-17F–deficient controls ( n =3–4), nephritic wild-type ( n =5), and nephritic IL-17F–deficient mice ( n =4) at day 8 of NTN. (G) Real-time RT-PCR analyses of renal mRNA expression of different chemokines of controls ( n =6), nephritic wild-type ( n =11), and nephritic IL-17F–deficient mice ( n =11) 8 days after induction of nephritis. mRNA levels are expressed as x-fold of controls. Symbols represent individual data points with the mean as horizontal line, or bar graphs with the mean±SD. * P
    Figure Legend Snippet: IL-17F drives renal tissue injury by mediating neutrophil infiltration. (A) Representative photographs of PAS-stained kidney sections from control, nephritic wild-type, and nephritic IL-17F–deficient mice at day 8 of NTN (original magnification ×400). (B) Quantification of glomerular crescent formation, interstitial area, and glomerular sclerosis of controls ( n =6), nephritic wild-type ( n =11), and nephritic IL-17F–deficient mice ( n =11) 8 days after induction of nephritis. (C) BUN levels, serum creatinine, and ACR of the aforementioned groups 8 days after induction of nephritis. (D) Quantification of tubulointerstitial GR-1 + cells, F4/80 + cells, and glomerular MAC-2 + cells in the aforementioned groups 8 days after induction of nephritis. (E) Representative FACS plots showing renal neutrophils (defined as CD45 + CD11b + Ly6G + cells) in nephritic wild-type and nephritic IL-17F–deficient mice and (F) quantification of FACS analysis of renal and splenic neutrophils of wild-type controls ( n =3–4), IL-17F–deficient controls ( n =3–4), nephritic wild-type ( n =5), and nephritic IL-17F–deficient mice ( n =4) at day 8 of NTN. (G) Real-time RT-PCR analyses of renal mRNA expression of different chemokines of controls ( n =6), nephritic wild-type ( n =11), and nephritic IL-17F–deficient mice ( n =11) 8 days after induction of nephritis. mRNA levels are expressed as x-fold of controls. Symbols represent individual data points with the mean as horizontal line, or bar graphs with the mean±SD. * P

    Techniques Used: Staining, Mouse Assay, FACS, Quantitative RT-PCR, Expressing

    IL-17F induces CXCL1 and CXCL5 expression in resident kidney cells. (A, B) Real-time RT-PCR analyses of Cxcl1 and Cxcl5 mRNA expression from (A) murine tubular cells and (B) murine mesangial cells after 4 hours of in vitro culture with medium alone or with addition of IL-17F (100 ng/ml), TNF α (10 ng/ml), or the combination of IL-17F (100 ng/ml) and TNF α (10 ng/ml). (C, D) ELISA analyses of CXCL1 and CXCL5 protein levels from supernatants of cultured murine tubular and murine mesangial cells after 24 hours of incubation under the conditions mentioned above. mRNA levels are expressed as x-fold of controls (= medium alone condition). n =3–4 mice per condition, data are presented as bar graphs with the mean±SD. * P
    Figure Legend Snippet: IL-17F induces CXCL1 and CXCL5 expression in resident kidney cells. (A, B) Real-time RT-PCR analyses of Cxcl1 and Cxcl5 mRNA expression from (A) murine tubular cells and (B) murine mesangial cells after 4 hours of in vitro culture with medium alone or with addition of IL-17F (100 ng/ml), TNF α (10 ng/ml), or the combination of IL-17F (100 ng/ml) and TNF α (10 ng/ml). (C, D) ELISA analyses of CXCL1 and CXCL5 protein levels from supernatants of cultured murine tubular and murine mesangial cells after 24 hours of incubation under the conditions mentioned above. mRNA levels are expressed as x-fold of controls (= medium alone condition). n =3–4 mice per condition, data are presented as bar graphs with the mean±SD. * P

    Techniques Used: Expressing, Quantitative RT-PCR, In Vitro, Enzyme-linked Immunosorbent Assay, Cell Culture, Incubation, Mouse Assay

    IL-17F serves a nonredundant function in crescentic GN. (A) Representative FACS plots showing IL-17A and IL-17F expression after PMA/ionomycin stimulation by CD4 + T cells and γδ T cells isolated from kidneys of nephritic IL-17A–deficient mice and nephritic IL-17F–deficient mice pregated for live CD45 + cells on day 8 of NTN. (B) Quantification of renal CD4 + T cells and γδ T cells of nephritic IL-17A–deficient mice ( n =6) and nephritic IL-17F–deficient mice ( n =6) at day 8 of NTN. (C) Representative photographs of PAS-stained kidney sections from nephritic IL-17A–deficient mice and nephritic IL-17F–deficient mice at day 8 of NTN (original magnification ×400). (D) Quantification of glomerular crescent formation, interstitial area, glomerular sclerosis, BUN levels, and ACR of nephritic IL-17A–deficient mice ( n =6) and nephritic IL-17F–deficient mice ( n =5) at day 8 of NTN. Symbols represent individual data points with the mean as horizontal line. * P
    Figure Legend Snippet: IL-17F serves a nonredundant function in crescentic GN. (A) Representative FACS plots showing IL-17A and IL-17F expression after PMA/ionomycin stimulation by CD4 + T cells and γδ T cells isolated from kidneys of nephritic IL-17A–deficient mice and nephritic IL-17F–deficient mice pregated for live CD45 + cells on day 8 of NTN. (B) Quantification of renal CD4 + T cells and γδ T cells of nephritic IL-17A–deficient mice ( n =6) and nephritic IL-17F–deficient mice ( n =6) at day 8 of NTN. (C) Representative photographs of PAS-stained kidney sections from nephritic IL-17A–deficient mice and nephritic IL-17F–deficient mice at day 8 of NTN (original magnification ×400). (D) Quantification of glomerular crescent formation, interstitial area, glomerular sclerosis, BUN levels, and ACR of nephritic IL-17A–deficient mice ( n =6) and nephritic IL-17F–deficient mice ( n =5) at day 8 of NTN. Symbols represent individual data points with the mean as horizontal line. * P

    Techniques Used: FACS, Expressing, Isolation, Mouse Assay, Staining

    IL-17F neutralization attenuates crescentic GN. (A) Representative photographs of PAS-stained kidney sections from control mice, wild-type mice treated with isotype antibody (250 μ g intraperitoneal injection on day 0 and day 4 of NTN), and wild-type mice treated with anti–IL-17F antibody (250 μ g intraperitoneal injection on day 0 and day 4 of NTN), at day 8 of NTN (original magnification ×400). (B) Quantification of glomerular crescent formation, interstitial area, and glomerular sclerosis of controls ( n =3), wild-type mice treated with isotype antibody ( n =9), and wild-type mice treated with anti–IL-17F antibody ( n =9) 8 days after induction of nephritis. (C) BUN levels, serum creatinine, and ACR of the aforementioned groups 8 days after induction of nephritis. (D) Quantification of tubulointerstitial GR-1 + cells, tubulointerstitial F4/80 + cells, and glomerular MAC-2 + cells of the aforementioned groups 8 days after induction of nephritis. (E) Quantification of tubulointerstitial CD3 + T cells and glomerular CD3 + T cells of the aforementioned groups 8 days after induction of nephritis. (F) Real-time RT-PCR analyses of renal mRNA expression of different chemokines in the aforementioned groups. mRNA levels are expressed as x-fold of controls. Symbols represent individual data points with the mean as horizontal line, or bar graphs with the mean±SD. * P
    Figure Legend Snippet: IL-17F neutralization attenuates crescentic GN. (A) Representative photographs of PAS-stained kidney sections from control mice, wild-type mice treated with isotype antibody (250 μ g intraperitoneal injection on day 0 and day 4 of NTN), and wild-type mice treated with anti–IL-17F antibody (250 μ g intraperitoneal injection on day 0 and day 4 of NTN), at day 8 of NTN (original magnification ×400). (B) Quantification of glomerular crescent formation, interstitial area, and glomerular sclerosis of controls ( n =3), wild-type mice treated with isotype antibody ( n =9), and wild-type mice treated with anti–IL-17F antibody ( n =9) 8 days after induction of nephritis. (C) BUN levels, serum creatinine, and ACR of the aforementioned groups 8 days after induction of nephritis. (D) Quantification of tubulointerstitial GR-1 + cells, tubulointerstitial F4/80 + cells, and glomerular MAC-2 + cells of the aforementioned groups 8 days after induction of nephritis. (E) Quantification of tubulointerstitial CD3 + T cells and glomerular CD3 + T cells of the aforementioned groups 8 days after induction of nephritis. (F) Real-time RT-PCR analyses of renal mRNA expression of different chemokines in the aforementioned groups. mRNA levels are expressed as x-fold of controls. Symbols represent individual data points with the mean as horizontal line, or bar graphs with the mean±SD. * P

    Techniques Used: Neutralization, Staining, Mouse Assay, Injection, Quantitative RT-PCR, Expressing

    IL-17F–producing CD4 + T cells aggravate renal tissue injury in Rag1 −/− mice. (A) Representative photographs of PAS-stained kidney sections at day 8 of NTN from RAG1 −/− control mice, nephritic RAG1 −/− mice repopulated with 1.5×10 6 wild-type CD4 + T cells, and nephritic RAG1 −/− mice repopulated with 1.5×10 6 IL-17F −/− CD4 + T cells (original magnification ×400). (B) Quantification of glomerular crescent formation, interstitial area, and glomerular sclerosis at day 8 of NTN of RAG1 −/− controls ( n =2), nephritic RAG1 −/− mice repopulated with wild-type CD4 + T cells ( n =7), and nephritic RAG1 −/− mice repopulated with IL-17F −/− CD4 + T cells ( n =8). (C) BUN levels, serum creatinine, and ACR of the aforementioned groups 8 days after induction of nephritis. (D) Quantification of tubulointerstitial GR-1 + cells, tubulointerstitial F4/80 + cells, and glomerular MAC-2 + cells of the aforementioned groups 8 days after induction of nephritis. Symbols represent individual data points with the mean as horizontal line. * P
    Figure Legend Snippet: IL-17F–producing CD4 + T cells aggravate renal tissue injury in Rag1 −/− mice. (A) Representative photographs of PAS-stained kidney sections at day 8 of NTN from RAG1 −/− control mice, nephritic RAG1 −/− mice repopulated with 1.5×10 6 wild-type CD4 + T cells, and nephritic RAG1 −/− mice repopulated with 1.5×10 6 IL-17F −/− CD4 + T cells (original magnification ×400). (B) Quantification of glomerular crescent formation, interstitial area, and glomerular sclerosis at day 8 of NTN of RAG1 −/− controls ( n =2), nephritic RAG1 −/− mice repopulated with wild-type CD4 + T cells ( n =7), and nephritic RAG1 −/− mice repopulated with IL-17F −/− CD4 + T cells ( n =8). (C) BUN levels, serum creatinine, and ACR of the aforementioned groups 8 days after induction of nephritis. (D) Quantification of tubulointerstitial GR-1 + cells, tubulointerstitial F4/80 + cells, and glomerular MAC-2 + cells of the aforementioned groups 8 days after induction of nephritis. Symbols represent individual data points with the mean as horizontal line. * P

    Techniques Used: Mouse Assay, Staining

    IL-17F expression in experimental GN. (A) Representative FACS plots showing IL-17F expression after PMA/ionomycin stimulation by selected cell subsets pregated for live CD45 + cells in kidney (above) and spleen (below) on days 0 (control) and 3 of NTN. (B) Quantification of leukocyte subset contribution to renal (left) and splenic (right) IL-17F production in the course of NTN at indicated time points. (C, D) Quantification of IL-17F production from (C) renal CD4 + T cells and γδ T cells, as well as from (D) splenic CD4 + T cells and γδ T cells in the course of NTN at indicated time points. (E, F) Quantification of (E) IL-17F serum levels and (F) IL-17F in supernatants of spleen cells restimulated with sIgG at indicated time points in the course of NTN. (G) Representative FACS plots showing IFN γ , IL-17F, and IL-17A expression by CD4 + T cells and γδ T cells isolated from kidneys of nephritic wild-type mice pregated for live CD45 + cells after PMA/ionomycin stimulation on day 8 of NTN. n =3–4 mice per time point, symbols represent individual data points with the mean as horizontal line, or bar graphs with the mean±SD, * P
    Figure Legend Snippet: IL-17F expression in experimental GN. (A) Representative FACS plots showing IL-17F expression after PMA/ionomycin stimulation by selected cell subsets pregated for live CD45 + cells in kidney (above) and spleen (below) on days 0 (control) and 3 of NTN. (B) Quantification of leukocyte subset contribution to renal (left) and splenic (right) IL-17F production in the course of NTN at indicated time points. (C, D) Quantification of IL-17F production from (C) renal CD4 + T cells and γδ T cells, as well as from (D) splenic CD4 + T cells and γδ T cells in the course of NTN at indicated time points. (E, F) Quantification of (E) IL-17F serum levels and (F) IL-17F in supernatants of spleen cells restimulated with sIgG at indicated time points in the course of NTN. (G) Representative FACS plots showing IFN γ , IL-17F, and IL-17A expression by CD4 + T cells and γδ T cells isolated from kidneys of nephritic wild-type mice pregated for live CD45 + cells after PMA/ionomycin stimulation on day 8 of NTN. n =3–4 mice per time point, symbols represent individual data points with the mean as horizontal line, or bar graphs with the mean±SD, * P

    Techniques Used: Expressing, FACS, Isolation, Mouse Assay

    IL-17F promotes autoimmune disease in pristane-induced lupus nephritis. (A) Representative FACS plots showing IFN γ , IL-17F, and IL-17A expression after PMA/ionomycin stimulation by CD4 + T cells and γδ T cells isolated from kidneys of wild-type and IL-17F–deficient mice pregated for live CD45 + cells 11 months after disease induction. (B) Representative photographs of PAS-stained kidney sections from control mice, wild-type lupus mice, and IL-17F–deficient lupus mice 11 months after disease induction (original magnification ×400). (C) Quantification of glomerular abnormalities and glomerular area of controls ( n =5), wild-type lupus mice ( n =4), and IL-17F–deficient lupus mice ( n =8) 11 months after disease induction. (D) Quantification of immunohistochemically stained tubulointerstitial and glomerular GR-1 + cells, as well as quantification of FACS analysis of renal neutrophils (defined as CD45 + CD11b + Ly6G + cells) in the aforementioned groups 11 months after disease induction. (E) Kaplan–Meier plot of survival in control and lupus nephritis groups over the course of disease. (F) ACR of the groups mentioned above over the course of disease. (G, H) Representative photographs of (G) Ki-67- and (H) mouse IgG-stained kidney sections of the aforementioned groups 11 months after disease induction. (I) ELISA analyses of circulating mouse autoantibodies against dsDNA and U1-RNP from sera of control and lupus nephritis mice 11 months after disease induction. Symbols represent mean values±SD connected by lines, or individual data points with the mean as horizontal line. * P
    Figure Legend Snippet: IL-17F promotes autoimmune disease in pristane-induced lupus nephritis. (A) Representative FACS plots showing IFN γ , IL-17F, and IL-17A expression after PMA/ionomycin stimulation by CD4 + T cells and γδ T cells isolated from kidneys of wild-type and IL-17F–deficient mice pregated for live CD45 + cells 11 months after disease induction. (B) Representative photographs of PAS-stained kidney sections from control mice, wild-type lupus mice, and IL-17F–deficient lupus mice 11 months after disease induction (original magnification ×400). (C) Quantification of glomerular abnormalities and glomerular area of controls ( n =5), wild-type lupus mice ( n =4), and IL-17F–deficient lupus mice ( n =8) 11 months after disease induction. (D) Quantification of immunohistochemically stained tubulointerstitial and glomerular GR-1 + cells, as well as quantification of FACS analysis of renal neutrophils (defined as CD45 + CD11b + Ly6G + cells) in the aforementioned groups 11 months after disease induction. (E) Kaplan–Meier plot of survival in control and lupus nephritis groups over the course of disease. (F) ACR of the groups mentioned above over the course of disease. (G, H) Representative photographs of (G) Ki-67- and (H) mouse IgG-stained kidney sections of the aforementioned groups 11 months after disease induction. (I) ELISA analyses of circulating mouse autoantibodies against dsDNA and U1-RNP from sera of control and lupus nephritis mice 11 months after disease induction. Symbols represent mean values±SD connected by lines, or individual data points with the mean as horizontal line. * P

    Techniques Used: FACS, Expressing, Isolation, Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay

    6) Product Images from "Th17 cells enhance viral persistence and inhibit T cell cytotoxicity in a model of chronic virus infection"

    Article Title: Th17 cells enhance viral persistence and inhibit T cell cytotoxicity in a model of chronic virus infection

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20082030

    IL-17 inhibits antiviral cytotoxic T cell function. (A) Splenocytes from B6 mice at 8 d after infection were pretreated for 24 h or were simultaneously treated with IL-17 (IL-17 Pre or IL-17 Sim, respectively), and were then subjected to a 51 Cr-release assay using VP2 121-130 –loaded EL-4 target cells. (B) Splenocytes (effector cells) from TMEV-infected B6 mice were co-cultured with naive B6 splenocytes (target cells) loaded with VP2 121-130 for 24 h in the presence of IL-17 at an E/T ratio of 10:1. The numbers in the plots indicate the frequency of granzyme B– or IFN-γ–positive CD8 + T cells. (C) Splenocytes from naive B6 mice were loaded with VP2 121-130 or OVA 323-339 peptides and labeled with a lower or higher concentration of CFSE, respectively, as target cells. The labeled target and effector cells were pretreated separately with IL-17 for 24 h and co-cultured or cultured together in the presence of IL-17F or IL-17 for an additional 60 h. The numbers in each histogram represent percentages of the lower (VP2 121-130 ) and higher (OVA 323-339 ) concentrations of CFSE-labeled cells. The arrow shows the specific killing of target cells. (D) Apoptosis levels of effector and target cells from the cultures in C at an E/T ratio of 10:1 were assessed after a 24-h incubation (percentages are shown). (E) Levels of target cell destruction in the presence of Fas:Fc were assessed 60 h after co-culture with effector cells at an E/T ratio of 30:1 (percentages are shown). The arrows show virus-specific killing. (F) Levels of antigen-loaded B cell killing by activated CD4 + T cells were determined. Peptide-loaded B cells were co-cultured in the presence of LPS and PBS or IL-17 at a 1:5 ratio for 24 h with effector OT-II T cells that were preactivated with anti-CD3/CD28 antibodies. The numbers in each plot indicate the frequency of Annexin V + B220 + B cells. (G) Intracellular expression levels of Bcl-xl and Bcl-2 proteins in LPS-stimulated B cells, BM cells, and BMDCs from naive B6 mice were assessed after a 24-h incubation with PBS (shaded histogram) or IL-17 (open histogram). Relative median fluorescence intensity differences between the IL-17 and the PBS group are illustrated on the flow histograms. Data are representative of four independent experiments.
    Figure Legend Snippet: IL-17 inhibits antiviral cytotoxic T cell function. (A) Splenocytes from B6 mice at 8 d after infection were pretreated for 24 h or were simultaneously treated with IL-17 (IL-17 Pre or IL-17 Sim, respectively), and were then subjected to a 51 Cr-release assay using VP2 121-130 –loaded EL-4 target cells. (B) Splenocytes (effector cells) from TMEV-infected B6 mice were co-cultured with naive B6 splenocytes (target cells) loaded with VP2 121-130 for 24 h in the presence of IL-17 at an E/T ratio of 10:1. The numbers in the plots indicate the frequency of granzyme B– or IFN-γ–positive CD8 + T cells. (C) Splenocytes from naive B6 mice were loaded with VP2 121-130 or OVA 323-339 peptides and labeled with a lower or higher concentration of CFSE, respectively, as target cells. The labeled target and effector cells were pretreated separately with IL-17 for 24 h and co-cultured or cultured together in the presence of IL-17F or IL-17 for an additional 60 h. The numbers in each histogram represent percentages of the lower (VP2 121-130 ) and higher (OVA 323-339 ) concentrations of CFSE-labeled cells. The arrow shows the specific killing of target cells. (D) Apoptosis levels of effector and target cells from the cultures in C at an E/T ratio of 10:1 were assessed after a 24-h incubation (percentages are shown). (E) Levels of target cell destruction in the presence of Fas:Fc were assessed 60 h after co-culture with effector cells at an E/T ratio of 30:1 (percentages are shown). The arrows show virus-specific killing. (F) Levels of antigen-loaded B cell killing by activated CD4 + T cells were determined. Peptide-loaded B cells were co-cultured in the presence of LPS and PBS or IL-17 at a 1:5 ratio for 24 h with effector OT-II T cells that were preactivated with anti-CD3/CD28 antibodies. The numbers in each plot indicate the frequency of Annexin V + B220 + B cells. (G) Intracellular expression levels of Bcl-xl and Bcl-2 proteins in LPS-stimulated B cells, BM cells, and BMDCs from naive B6 mice were assessed after a 24-h incubation with PBS (shaded histogram) or IL-17 (open histogram). Relative median fluorescence intensity differences between the IL-17 and the PBS group are illustrated on the flow histograms. Data are representative of four independent experiments.

    Techniques Used: Cell Function Assay, Mouse Assay, Infection, Release Assay, Cell Culture, Labeling, Concentration Assay, Incubation, Co-Culture Assay, Expressing, Fluorescence, Flow Cytometry

    7) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    8) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    9) Product Images from "Delineation of the Innate and Adaptive T-Cell Immune Outcome in the Human Host in Response to Campylobacter jejuni Infection"

    Article Title: Delineation of the Innate and Adaptive T-Cell Immune Outcome in the Human Host in Response to Campylobacter jejuni Infection

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0015398

    IL-17A and IL-17F reduce C . jejuni 11168H intracellular survival in intestinal epithelia. Confluent Caco-2 cells were exposed to individual cytokines for 24 hours prior to infection with C. jejuni 11168H WT strain (MOI = 100) for 3 hours at 37°C. Cell lysates were serially diluted and plated for total viable bacterial counts (adhesion + invasion). In parallel, another set of infected cells was exposed to 150 µg/ml gentamicin for 2 hours (to kill extracellular adhered bacteria) and lysates plated for enumeration of viable intracellular bacteria. Data represent average percentage cfu obtained in treated versus untreated cells (the latter set at 100%). Statistical analysis of 3 independent experiments performed in duplicate is shown versus untreated cells (Data shown as the median + range).
    Figure Legend Snippet: IL-17A and IL-17F reduce C . jejuni 11168H intracellular survival in intestinal epithelia. Confluent Caco-2 cells were exposed to individual cytokines for 24 hours prior to infection with C. jejuni 11168H WT strain (MOI = 100) for 3 hours at 37°C. Cell lysates were serially diluted and plated for total viable bacterial counts (adhesion + invasion). In parallel, another set of infected cells was exposed to 150 µg/ml gentamicin for 2 hours (to kill extracellular adhered bacteria) and lysates plated for enumeration of viable intracellular bacteria. Data represent average percentage cfu obtained in treated versus untreated cells (the latter set at 100%). Statistical analysis of 3 independent experiments performed in duplicate is shown versus untreated cells (Data shown as the median + range).

    Techniques Used: Infection

    10) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    11) Product Images from "Th17 cell-derived IL-17 is dispensable for B cell antibody production"

    Article Title: Th17 cell-derived IL-17 is dispensable for B cell antibody production

    Journal: Cytokine

    doi: 10.1016/j.cyto.2012.03.018

    Effect of IL-25 and IL-17F on B-cell Ig production Splenic B cells (2 × 10 6 cells/ml) were stimulated with 100 ng/ml of rmIL-17, rmIL-17F or rmIL-25 in the presence and absence of anti-CD40 mAb for 6 days. The levels of IgA, IgE, IgG1, IgG2a, IgG2b, IgG2c and IgG3 in the culture supernatants were measured by ELISA. Data show the mean ± SEM (n = 6). *p
    Figure Legend Snippet: Effect of IL-25 and IL-17F on B-cell Ig production Splenic B cells (2 × 10 6 cells/ml) were stimulated with 100 ng/ml of rmIL-17, rmIL-17F or rmIL-25 in the presence and absence of anti-CD40 mAb for 6 days. The levels of IgA, IgE, IgG1, IgG2a, IgG2b, IgG2c and IgG3 in the culture supernatants were measured by ELISA. Data show the mean ± SEM (n = 6). *p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    12) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    13) Product Images from "Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth"

    Article Title: Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth

    Journal: Oncogene

    doi: 10.1038/onc.2014.286

    IL-17A, IL-22, TNF-α and IL-6 contribute to TIL-derived supernatant (TIL SN)-mediated STAT3/NF-kB activation and mitogenic effect in CRC cells. ( a ) Representative western blotting showing p-STAT3 Tyr705, STAT3, p-NF-kB/p65 Ser536 and NF-kB/p65 expression in DLD-1 and HT-29 cells stimulated or not with IL-17A, IL-17F, IL-21, IL-22, TNF-α- and IL-6 (all used at 25 ng/ml) for 15 min. β-Actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. ( b ) Representative western blotting showing p-STAT3 Tyr705, STAT3, p-p65 Ser536 and NF-kB/p65 expression in DLD-1 and HT-29 cells stimulated or not with TIL SNs in the presence or absence of anti-IL-17A, anti-IL-17F, anti-IL-21, anti-IL-22, anti-TNF-α and anti-IL-6 (all used at 10 μg/ml) as indicated. β-Actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. ( c ) Representative histograms showing cell proliferation of DLD-1 and HT-29 cells stimulated as indicated in ( b ). Data indicate mean±s.e.m. of four experiments. Differences between groups were compared using one-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test. ( d ) Representative western blotting showing p-STAT3 Tyr705, STAT3, p-NF-kB/p65 Ser536 and NF-kB/p65 expression in DLD-1 and HT-29 cells stimulated or not with TIL SNs in the presence or absence of anti-IL-17A, anti-IL-22, anti-TNF-α and anti-IL-6, used in combination as indicated. β-Actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. ( e ) Representative histograms showing cell proliferation of DLD-1 and HT-29 cells stimulated as indicated in ( d ). Data indicate mean±s.e.m. of five experiments. Differences between groups were compared using one-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test. DLD-1: TIL SN+anti-IL-22+anti-IL-6- vs TIL SN-treated cells, * P
    Figure Legend Snippet: IL-17A, IL-22, TNF-α and IL-6 contribute to TIL-derived supernatant (TIL SN)-mediated STAT3/NF-kB activation and mitogenic effect in CRC cells. ( a ) Representative western blotting showing p-STAT3 Tyr705, STAT3, p-NF-kB/p65 Ser536 and NF-kB/p65 expression in DLD-1 and HT-29 cells stimulated or not with IL-17A, IL-17F, IL-21, IL-22, TNF-α- and IL-6 (all used at 25 ng/ml) for 15 min. β-Actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. ( b ) Representative western blotting showing p-STAT3 Tyr705, STAT3, p-p65 Ser536 and NF-kB/p65 expression in DLD-1 and HT-29 cells stimulated or not with TIL SNs in the presence or absence of anti-IL-17A, anti-IL-17F, anti-IL-21, anti-IL-22, anti-TNF-α and anti-IL-6 (all used at 10 μg/ml) as indicated. β-Actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. ( c ) Representative histograms showing cell proliferation of DLD-1 and HT-29 cells stimulated as indicated in ( b ). Data indicate mean±s.e.m. of four experiments. Differences between groups were compared using one-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test. ( d ) Representative western blotting showing p-STAT3 Tyr705, STAT3, p-NF-kB/p65 Ser536 and NF-kB/p65 expression in DLD-1 and HT-29 cells stimulated or not with TIL SNs in the presence or absence of anti-IL-17A, anti-IL-22, anti-TNF-α and anti-IL-6, used in combination as indicated. β-Actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. ( e ) Representative histograms showing cell proliferation of DLD-1 and HT-29 cells stimulated as indicated in ( d ). Data indicate mean±s.e.m. of five experiments. Differences between groups were compared using one-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test. DLD-1: TIL SN+anti-IL-22+anti-IL-6- vs TIL SN-treated cells, * P

    Techniques Used: Derivative Assay, Activation Assay, Western Blot, Expressing

    The neoplastic area of CRC samples is massively infiltrated with Th17-related cytokine-, TNF-α- and IL-6-producing cells. ( a ) IFN-γ, IL-17A, IL-17F, IL-21, IL-22, TNF-α and IL-6 proteins were analyzed by enzyme-linked immunosorbent assay (ELISA) in LPMC-derived supernatants (LPMC SNs) and TIL-derived supernatants (TL SNs), and data are expressed as pg/ml supernatants. Data indicate mean±s.e.m. of six experiments. Differences were calculated using the two-tailed Student's t -test. ( b ) Characterization of immune cell infiltrate in LPMCs and TIsL. Representative histograms showing the fraction of T cells, natural killer T (NKT) cells, NK cells, macrophages and B cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. CD45+ cells were gated and analyzed for the indicated markers. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test. ( c ) Representative histograms showing the fraction of IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-expressing CD45+ cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test.
    Figure Legend Snippet: The neoplastic area of CRC samples is massively infiltrated with Th17-related cytokine-, TNF-α- and IL-6-producing cells. ( a ) IFN-γ, IL-17A, IL-17F, IL-21, IL-22, TNF-α and IL-6 proteins were analyzed by enzyme-linked immunosorbent assay (ELISA) in LPMC-derived supernatants (LPMC SNs) and TIL-derived supernatants (TL SNs), and data are expressed as pg/ml supernatants. Data indicate mean±s.e.m. of six experiments. Differences were calculated using the two-tailed Student's t -test. ( b ) Characterization of immune cell infiltrate in LPMCs and TIsL. Representative histograms showing the fraction of T cells, natural killer T (NKT) cells, NK cells, macrophages and B cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. CD45+ cells were gated and analyzed for the indicated markers. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test. ( c ) Representative histograms showing the fraction of IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-expressing CD45+ cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Derivative Assay, Two Tailed Test, Isolation, Expressing

    Analysis of cytokine production and T-bet/RORγt expression in LPMC- and TIL-derived CD3+CD8− subsets. ( a ) Representative histograms showing the fraction of IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-expressing CD3+CD8− cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test. Right insets. Representative dot plots showing the percentage of IFN-γ- and/or IL-21-, IL-17A- and/or IL-17F-, IL-22- and/or IL-6- and TNF-α-producing CD3+CD8− cells in LPMCs and TILs. The numbers indicate the percentage of cells in the designated quadrants. ( b ) Representative histograms showing the fraction of T-bet+ and/or Rorγt+ CD3+CD8− cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test. Right insets. Representative dot plots showing the percentage of T-bet+ and/or Rorγt+ CD3+CD8− cells in LPMCs and TILs. The numbers indicate the percentage of cells in the designated quadrants. Staining of LPMCs with APC- and PE-Cy7-conjugated control isotype IgG is also shown. ( c ) Representative dot plots showing the ability to produce IFN-γ and/or IL-17A by the indicated subsets of TILs. The numbers indicate the percentage of cells in the designated quadrants. ( d ) Representative histograms showing the percentage of IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-producing CD3+CD8− cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of one patient undergoing colectomy for sporadic CRC . IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-producing CD3+CD8− cells were gated and analyzed for the indicated markers. The example is representative of 10 independent experiments in which cells isolated from 10 patients undergoing colectomy for sporadic CRC were analyzed.
    Figure Legend Snippet: Analysis of cytokine production and T-bet/RORγt expression in LPMC- and TIL-derived CD3+CD8− subsets. ( a ) Representative histograms showing the fraction of IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-expressing CD3+CD8− cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test. Right insets. Representative dot plots showing the percentage of IFN-γ- and/or IL-21-, IL-17A- and/or IL-17F-, IL-22- and/or IL-6- and TNF-α-producing CD3+CD8− cells in LPMCs and TILs. The numbers indicate the percentage of cells in the designated quadrants. ( b ) Representative histograms showing the fraction of T-bet+ and/or Rorγt+ CD3+CD8− cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of 14 patients undergoing colectomy for sporadic CRC. Data are expressed as mean±s.e.m. and differences were calculated using the two-tailed Student's t -test. Right insets. Representative dot plots showing the percentage of T-bet+ and/or Rorγt+ CD3+CD8− cells in LPMCs and TILs. The numbers indicate the percentage of cells in the designated quadrants. Staining of LPMCs with APC- and PE-Cy7-conjugated control isotype IgG is also shown. ( c ) Representative dot plots showing the ability to produce IFN-γ and/or IL-17A by the indicated subsets of TILs. The numbers indicate the percentage of cells in the designated quadrants. ( d ) Representative histograms showing the percentage of IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-producing CD3+CD8− cells in LPMCs and TILs isolated from adjacent tumor and non-tumor areas of one patient undergoing colectomy for sporadic CRC . IFN-γ-, IL-17A-, IL-17F-, IL-21-, IL-22-, TNF-α- and IL-6-producing CD3+CD8− cells were gated and analyzed for the indicated markers. The example is representative of 10 independent experiments in which cells isolated from 10 patients undergoing colectomy for sporadic CRC were analyzed.

    Techniques Used: Expressing, Derivative Assay, Isolation, Two Tailed Test, Staining

    Reduced STAT3/NF-kB activation and reduced expression of IL-17A, IL-21, IL-22, TNF-α and IL-6 are seen in the colonic tumors of BP-1-102-treated Apc min/+ mice. ( a ) Representative images showing p-STAT3 Tyr705- or p-NF-kB/p65 Ser536-positive cells in colonic sections taken from the tumor areas Apc min/+ mice treated with either dimethyl sulfoxide (DMSO) (CTR) or BP-1-102 and killed on day 56. The scale bars are 20 μm. One of six representative experiments in which similar results were obtained is shown. Right insets. Quantification of p-STAT3 Tyr705- or p-NF-kB/p65 Ser536-positive infiltrating and epithelial cells in colonic sections taken from the tumor areas of Apc min/+ mice treated with either DMSO (CTR) or BP-1-102 and killed on day 56. Data are presented as mean values of positive cells per high power field (h.p.f.)±s.e.m. of three independent experiments in which two sections per group were analyzed. Differences were calculated using the two-tailed Student's t -test. NT, non-tumor area; T, tumor area. ( b ) IFN-γ, IL-17A, IL-17F, IL-21, IL-22, TNF-α- and IL-6 expression was assessed by real-time PCR in colonic tissues taken from Apc min/+ mice treated with either DMSO (CTR) or BP-1-102 and killed on day 56. Values are mean±s.e.m. of two independent experiments containing at least three mice per group. Differences were calculated using the two-tailed Student's t -test. ND, not detectable; NT, non-tumor area; T, tumor area.
    Figure Legend Snippet: Reduced STAT3/NF-kB activation and reduced expression of IL-17A, IL-21, IL-22, TNF-α and IL-6 are seen in the colonic tumors of BP-1-102-treated Apc min/+ mice. ( a ) Representative images showing p-STAT3 Tyr705- or p-NF-kB/p65 Ser536-positive cells in colonic sections taken from the tumor areas Apc min/+ mice treated with either dimethyl sulfoxide (DMSO) (CTR) or BP-1-102 and killed on day 56. The scale bars are 20 μm. One of six representative experiments in which similar results were obtained is shown. Right insets. Quantification of p-STAT3 Tyr705- or p-NF-kB/p65 Ser536-positive infiltrating and epithelial cells in colonic sections taken from the tumor areas of Apc min/+ mice treated with either DMSO (CTR) or BP-1-102 and killed on day 56. Data are presented as mean values of positive cells per high power field (h.p.f.)±s.e.m. of three independent experiments in which two sections per group were analyzed. Differences were calculated using the two-tailed Student's t -test. NT, non-tumor area; T, tumor area. ( b ) IFN-γ, IL-17A, IL-17F, IL-21, IL-22, TNF-α- and IL-6 expression was assessed by real-time PCR in colonic tissues taken from Apc min/+ mice treated with either DMSO (CTR) or BP-1-102 and killed on day 56. Values are mean±s.e.m. of two independent experiments containing at least three mice per group. Differences were calculated using the two-tailed Student's t -test. ND, not detectable; NT, non-tumor area; T, tumor area.

    Techniques Used: Activation Assay, Expressing, Mouse Assay, Two Tailed Test, Real-time Polymerase Chain Reaction

    14) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    15) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    16) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    17) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    18) Product Images from "Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells"

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3600

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells
    Figure Legend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Techniques Used: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.
    Figure Legend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Figure Legend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Techniques Used: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

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    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Collectively, these data demonstrated that activation of HSCs was decreased in mice treated with Ad-ACVR2A shRNA. .. In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ).

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Collectively, these data demonstrated that activation of HSCs was decreased in mice treated with Ad-ACVR2A shRNA. .. Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ).

    Infection:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: .. Furthermore, in order to disrupt activin/ACVR2A signaling in primary mHSCs, cells were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with IL-17A or IL-17F for an additional 48 h. The results demonstrated that IL-17-induced overproduction of collagens was suppressed when ACVR2A was inhibited. .. In addition, the expression levels of α-SMA and p-Smad2 were decreased in mHSCs infected with Ad-ACVR2A shRNA ( ).

    shRNA:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: .. Furthermore, in order to disrupt activin/ACVR2A signaling in primary mHSCs, cells were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with IL-17A or IL-17F for an additional 48 h. The results demonstrated that IL-17-induced overproduction of collagens was suppressed when ACVR2A was inhibited. .. In addition, the expression levels of α-SMA and p-Smad2 were decreased in mHSCs infected with Ad-ACVR2A shRNA ( ).

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Collectively, these data demonstrated that activation of HSCs was decreased in mice treated with Ad-ACVR2A shRNA. .. Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Con A administration stimulates activation of activin and IL-17 signaling in the liver In the present study, liver and serum samples were collected from the control and chronic Con A-treated mice at the end of week 6 ( ). .. The contents of activin A, and IL-17A and IL-17F in the serum and liver tissue samples were detected using corresponding ELISA kits. .. Activin A levels were increased in Con A-treated mice compared with in the control mice ( ).

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ). ..

    Activation Assay:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Collectively, these data demonstrated that activation of HSCs was decreased in mice treated with Ad-ACVR2A shRNA. .. Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ).

    In Vitro:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Collectively, these data demonstrated that activation of HSCs was decreased in mice treated with Ad-ACVR2A shRNA. .. Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ).

    Produced:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro In order to determine the role of the activin/ACVR2A/Smad2 pathway in immune-associated liver fibrosis, primary mHSCs were isolated from normal liver samples and were treated with IL-17A and IL-17F ( ). .. Following stimulation with IL-17A or IL-17F, mHSCs produced more activin A, as evidenced by ELISA ( ). ..

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Notably, in addition to TGF-β, the present study demonstrated that mHSCs generated more activin A in response to IL-17 stimulation. .. Furthermore, mHSCs with reducedACVR2A expression produced less collagens in the presence of IL-17A or IL-17F; α-SMA expression was also decreased. .. Wada et al previously reported that exogenous activin A activates HSCs and increases their collagen production and α-SMA expression ( ).

    Expressing:

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells
    Article Snippet: Notably, in addition to TGF-β, the present study demonstrated that mHSCs generated more activin A in response to IL-17 stimulation. .. Furthermore, mHSCs with reducedACVR2A expression produced less collagens in the presence of IL-17A or IL-17F; α-SMA expression was also decreased. .. Wada et al previously reported that exogenous activin A activates HSCs and increases their collagen production and α-SMA expression ( ).

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    PeproTech il 17░a
    <t>IL-17░A</t> promoted the migration and invasion of NSCLC cells in vitro . The migration activities of A549 ( A ) and H460 ( B ) cells, and the invasion activities of A549 ( C ) and H460 ( D ) cells before and after treatment of rhIL-17░A was analyzed using transwell assay. One representative analysis is shown. Data are presented as a histogram. ( E) The expression of N-cadherin in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. ( F) The relative expression of N-cadherin in A549 and H460 cells before and after treatment with rhIL-17░A was analyzed using qPCR. (G) The expressions of IL-17░A and CD31 in tumor tissues from NSCLC patients were detected by immunofluorescence. * indicates P
    Il 17░A, supplied by PeproTech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 17░a/product/PeproTech
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 17░a - by Bioz Stars, 2021-05
    93/100 stars
      Buy from Supplier

    96
    PeproTech recombinant murine il 17a
    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant <t>IL-17A</t> or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.
    Recombinant Murine Il 17a, supplied by PeproTech, 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/recombinant murine il 17a/product/PeproTech
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant murine il 17a - by Bioz Stars, 2021-05
    96/100 stars
      Buy from Supplier

    N/A
    Murine IL 17 ELISA development kit contains the key components required for the quantitative measurement of natural and or recombinant murine IL 17 in a sandwich ELISA format Using the
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    Rabbit Anti Human IL 17B Source Polyclonal Rabbit Formulation Lyophilized Produced from sera of rabbits pre immunized with highly pure 98 recombinant hIL 17B Anti Human IL 17B specific antibody
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    IL-17░A promoted the migration and invasion of NSCLC cells in vitro . The migration activities of A549 ( A ) and H460 ( B ) cells, and the invasion activities of A549 ( C ) and H460 ( D ) cells before and after treatment of rhIL-17░A was analyzed using transwell assay. One representative analysis is shown. Data are presented as a histogram. ( E) The expression of N-cadherin in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. ( F) The relative expression of N-cadherin in A549 and H460 cells before and after treatment with rhIL-17░A was analyzed using qPCR. (G) The expressions of IL-17░A and CD31 in tumor tissues from NSCLC patients were detected by immunofluorescence. * indicates P

    Journal: Oncoimmunology

    Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

    doi: 10.1080/2162402X.2018.1461303

    Figure Lengend Snippet: IL-17░A promoted the migration and invasion of NSCLC cells in vitro . The migration activities of A549 ( A ) and H460 ( B ) cells, and the invasion activities of A549 ( C ) and H460 ( D ) cells before and after treatment of rhIL-17░A was analyzed using transwell assay. One representative analysis is shown. Data are presented as a histogram. ( E) The expression of N-cadherin in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. ( F) The relative expression of N-cadherin in A549 and H460 cells before and after treatment with rhIL-17░A was analyzed using qPCR. (G) The expressions of IL-17░A and CD31 in tumor tissues from NSCLC patients were detected by immunofluorescence. * indicates P

    Article Snippet: The proteins in lysates were resolved on SDS-PAGE gels, transferred onto PVDF membranes (Millipore, #ISEEQ00010), and immunoblotted with anti-human N-cadherin (Cell signaling Technology, #13116), snail (Cell signaling Technology, #3879), vimentin (Cell signaling Technology, #5741), p-STAT3 (Cell signaling Technology, #9145), p-p65 (Cell signaling Technology, #3033), cleaved-Notch1 (Cell signaling Technology, #4147), Oct4 (Cell signaling Technology, #2750), Nanog (Cell signaling Technology, #4903), Sox2 (Cell signaling Technology, #3579), IL-17░A (PeproTech, #500-P07) and anti-human β-actin antibody (ProteinTech, #20536-1-AP-50).

    Techniques: Migration, In Vitro, Transwell Assay, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence

    The level of Th17 cells was associated with poor survival in NSCLC patients. The relationship between Th17 frequency and TNM stage ( A ), tumor grade ( B ) and lymph node metastasis ( C ) of NSCLC patients, respectively. The changes of Th17 cell frequency in peripheral blood from NSCLC patients before and after chemotherapy with the status of PD ( D ), PR ( E ) and SD ( F ). ( G) IL-17░A expression in tumor tissues of NSCLC was analyzed by immunohistochemistry. ( H) Kaplan-Meier survival curves for NSCLC patients with lower and higher IL-17░A expression (immunohistochemistry analysis). *, ** and ns indicate P

    Journal: Oncoimmunology

    Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

    doi: 10.1080/2162402X.2018.1461303

    Figure Lengend Snippet: The level of Th17 cells was associated with poor survival in NSCLC patients. The relationship between Th17 frequency and TNM stage ( A ), tumor grade ( B ) and lymph node metastasis ( C ) of NSCLC patients, respectively. The changes of Th17 cell frequency in peripheral blood from NSCLC patients before and after chemotherapy with the status of PD ( D ), PR ( E ) and SD ( F ). ( G) IL-17░A expression in tumor tissues of NSCLC was analyzed by immunohistochemistry. ( H) Kaplan-Meier survival curves for NSCLC patients with lower and higher IL-17░A expression (immunohistochemistry analysis). *, ** and ns indicate P

    Article Snippet: The proteins in lysates were resolved on SDS-PAGE gels, transferred onto PVDF membranes (Millipore, #ISEEQ00010), and immunoblotted with anti-human N-cadherin (Cell signaling Technology, #13116), snail (Cell signaling Technology, #3879), vimentin (Cell signaling Technology, #5741), p-STAT3 (Cell signaling Technology, #9145), p-p65 (Cell signaling Technology, #3033), cleaved-Notch1 (Cell signaling Technology, #4147), Oct4 (Cell signaling Technology, #2750), Nanog (Cell signaling Technology, #4903), Sox2 (Cell signaling Technology, #3579), IL-17░A (PeproTech, #500-P07) and anti-human β-actin antibody (ProteinTech, #20536-1-AP-50).

    Techniques: Expressing, Immunohistochemistry

    IL-17░A promoted the CSC-like properties of NSCLC cells. A549 ( A ) and H460 ( B ) cells were cultured with rhIL-17░A for 7 days, and then collected for sphere assay. One representative photomicrograph is shown. Data are presented as a histogram. ( C) The expression of Oct4 in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. The apoptosis of A549 ( D ) and H460 ( E ) cells treated with or without docetaxel and rhIL-17░A was analyzed by flow cytometry. IL-17░A expression in IL-17░A-expressing and control vectors-tranfected H460 cells was analyzed by western blotting ( F ) and qPCR ( G ). ( H) IL-17░A promotes NSCLC tumor growth in vivo . IL-17░A expressing A549 cells were injected subcutaneously into nude mice (n = 6 mice/group). Tumor volumes were measured at 21 days after cell implantation. ( I) Lump images of xenograft tumors that were formed in nude mice at 21 days after cell implantation. ( J) The result of tumor weights at 21 days after cell implantation was shown as histogram. ( K) Tumor growth curve from 7 to 21 days after cell implantation was shown as statistical graph. * indicates P

    Journal: Oncoimmunology

    Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

    doi: 10.1080/2162402X.2018.1461303

    Figure Lengend Snippet: IL-17░A promoted the CSC-like properties of NSCLC cells. A549 ( A ) and H460 ( B ) cells were cultured with rhIL-17░A for 7 days, and then collected for sphere assay. One representative photomicrograph is shown. Data are presented as a histogram. ( C) The expression of Oct4 in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. The apoptosis of A549 ( D ) and H460 ( E ) cells treated with or without docetaxel and rhIL-17░A was analyzed by flow cytometry. IL-17░A expression in IL-17░A-expressing and control vectors-tranfected H460 cells was analyzed by western blotting ( F ) and qPCR ( G ). ( H) IL-17░A promotes NSCLC tumor growth in vivo . IL-17░A expressing A549 cells were injected subcutaneously into nude mice (n = 6 mice/group). Tumor volumes were measured at 21 days after cell implantation. ( I) Lump images of xenograft tumors that were formed in nude mice at 21 days after cell implantation. ( J) The result of tumor weights at 21 days after cell implantation was shown as histogram. ( K) Tumor growth curve from 7 to 21 days after cell implantation was shown as statistical graph. * indicates P

    Article Snippet: The proteins in lysates were resolved on SDS-PAGE gels, transferred onto PVDF membranes (Millipore, #ISEEQ00010), and immunoblotted with anti-human N-cadherin (Cell signaling Technology, #13116), snail (Cell signaling Technology, #3879), vimentin (Cell signaling Technology, #5741), p-STAT3 (Cell signaling Technology, #9145), p-p65 (Cell signaling Technology, #3033), cleaved-Notch1 (Cell signaling Technology, #4147), Oct4 (Cell signaling Technology, #2750), Nanog (Cell signaling Technology, #4903), Sox2 (Cell signaling Technology, #3579), IL-17░A (PeproTech, #500-P07) and anti-human β-actin antibody (ProteinTech, #20536-1-AP-50).

    Techniques: Cell Culture, Expressing, Western Blot, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction, In Vivo, Injection, Mouse Assay

    Blockade of STAT3/NF-κB/Notch1 signaling inhibited NSCLC stemness promoted by IL-17░A. (A) After the treatment with STAT3░or NF-κB or Notch1 inhibitor, the sphere forming of A549 and H460 cells induce by rhIL-17░A was blocked. One representative photomicrograph is shown. The results from A549 ( B ) and H460 ( C ) cells were shown as histogram. ( D) The expression of Oct4 in A549 and H460 cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by western blotting. The apoptosis of A549 ( E ) and H460 ( F ) cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by transwell assay. * indicates P

    Journal: Oncoimmunology

    Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

    doi: 10.1080/2162402X.2018.1461303

    Figure Lengend Snippet: Blockade of STAT3/NF-κB/Notch1 signaling inhibited NSCLC stemness promoted by IL-17░A. (A) After the treatment with STAT3░or NF-κB or Notch1 inhibitor, the sphere forming of A549 and H460 cells induce by rhIL-17░A was blocked. One representative photomicrograph is shown. The results from A549 ( B ) and H460 ( C ) cells were shown as histogram. ( D) The expression of Oct4 in A549 and H460 cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by western blotting. The apoptosis of A549 ( E ) and H460 ( F ) cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by transwell assay. * indicates P

    Article Snippet: The proteins in lysates were resolved on SDS-PAGE gels, transferred onto PVDF membranes (Millipore, #ISEEQ00010), and immunoblotted with anti-human N-cadherin (Cell signaling Technology, #13116), snail (Cell signaling Technology, #3879), vimentin (Cell signaling Technology, #5741), p-STAT3 (Cell signaling Technology, #9145), p-p65 (Cell signaling Technology, #3033), cleaved-Notch1 (Cell signaling Technology, #4147), Oct4 (Cell signaling Technology, #2750), Nanog (Cell signaling Technology, #4903), Sox2 (Cell signaling Technology, #3579), IL-17░A (PeproTech, #500-P07) and anti-human β-actin antibody (ProteinTech, #20536-1-AP-50).

    Techniques: Expressing, Western Blot, Transwell Assay

    The STAT3/NF-κB/Notch1 signaling was critical for IL-17░A-induced migration and invasion in NSCLC cells. (A) The activation of STAT3, NF-κB and Notch1in A549 and H460 cells treated with rhIL-17░A was analyzed using western blotting. ( B) The migration activities of A549 and H460 cells treated with or without rhIL-17░A and STAT3, or NF-κB, or Notch1 inhibitor were assessed by transwell assay. One representative analysis is shown. The data from A549 ( C ) and H460 ( D ) cells are presented as histogram. ( E) The invasion activities of A549 and H460 cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by transwell assay. One representative analysis is shown. The data from A549 ( F ) and H460 ( G ) cells are presented as a histogram. ( H) The expression of N-cadherin in A549 and H460 cells treated with or without rhIL-17░A and STAT3, or NF-κB, or Notch1 inhibitor was analyzed using western blotting. * indicates P

    Journal: Oncoimmunology

    Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

    doi: 10.1080/2162402X.2018.1461303

    Figure Lengend Snippet: The STAT3/NF-κB/Notch1 signaling was critical for IL-17░A-induced migration and invasion in NSCLC cells. (A) The activation of STAT3, NF-κB and Notch1in A549 and H460 cells treated with rhIL-17░A was analyzed using western blotting. ( B) The migration activities of A549 and H460 cells treated with or without rhIL-17░A and STAT3, or NF-κB, or Notch1 inhibitor were assessed by transwell assay. One representative analysis is shown. The data from A549 ( C ) and H460 ( D ) cells are presented as histogram. ( E) The invasion activities of A549 and H460 cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by transwell assay. One representative analysis is shown. The data from A549 ( F ) and H460 ( G ) cells are presented as a histogram. ( H) The expression of N-cadherin in A549 and H460 cells treated with or without rhIL-17░A and STAT3, or NF-κB, or Notch1 inhibitor was analyzed using western blotting. * indicates P

    Article Snippet: The proteins in lysates were resolved on SDS-PAGE gels, transferred onto PVDF membranes (Millipore, #ISEEQ00010), and immunoblotted with anti-human N-cadherin (Cell signaling Technology, #13116), snail (Cell signaling Technology, #3879), vimentin (Cell signaling Technology, #5741), p-STAT3 (Cell signaling Technology, #9145), p-p65 (Cell signaling Technology, #3033), cleaved-Notch1 (Cell signaling Technology, #4147), Oct4 (Cell signaling Technology, #2750), Nanog (Cell signaling Technology, #4903), Sox2 (Cell signaling Technology, #3579), IL-17░A (PeproTech, #500-P07) and anti-human β-actin antibody (ProteinTech, #20536-1-AP-50).

    Techniques: Migration, Activation Assay, Western Blot, Transwell Assay, Expressing

    Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Journal: International Journal of Molecular Medicine

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    doi: 10.3892/ijmm.2018.3600

    Figure Lengend Snippet: Ad-ACVR2A shRNA suppresses IL-17-induced activation of primary mHSCs in vitro . The primary mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA for 24 h, and were then stimulated with recombinant IL-17A or IL-17F for an additional 48 h. (A) Knockdown efficiency of Ad-ACVR2A shRNA was confirmed in normal mHSCs by western blot analysis. (B and D) Production of collagens I and VI in cell supernatants were determined by ELISA kits. (C and E) Protein expression levels of α-SMA, p-Smad2 and total Smad2 were determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells; NC, negative control; p, phosphorylated; shRNA, short hairpin RNA.

    Article Snippet: Recombinant murine IL-17A and IL-17F (PeproTech, Inc., Rocky Hill, NJ, USA) at 10, 30 or 100 ng/ml were used to stimulate mHSCs for 1, 3, 6, 12, 24 or 48 h. For some experiments, the mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA (multiplicity of infection, both 50) for 24 h, and were then stimulated with recombinant IL-17A (30 ng/ml) or IL-17F (30 ng/ml) for additional 48 h.

    Techniques: shRNA, Activation Assay, In Vitro, Infection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Negative Control

    IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Journal: International Journal of Molecular Medicine

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    doi: 10.3892/ijmm.2018.3600

    Figure Lengend Snippet: IL-17 induces activation of primary mHSCs in vitro . Recombinant mouse (A and B) IL-17A and (E and F) IL-17F (10, 30 or 100 ng/ml) were used to stimulate primary mHSCs for 48 h. In addition, mHSCs were treated with 30 ng/ml (C and D) IL-17A and (G and H) IL-17F for the indicated time periods. Activin A content in cell supernatants was determined by ELISA, and the expression levels of the marker for the activated HSCs, α-SMA, was determined by western blot analysis. Data are expressed as the means ± standard deviation (n=3/group). α-SMA, α-smooth muscle actin; IL-17, interleukin-17; mHSCs, mouse hepatic stellate cells

    Article Snippet: Recombinant murine IL-17A and IL-17F (PeproTech, Inc., Rocky Hill, NJ, USA) at 10, 30 or 100 ng/ml were used to stimulate mHSCs for 1, 3, 6, 12, 24 or 48 h. For some experiments, the mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA (multiplicity of infection, both 50) for 24 h, and were then stimulated with recombinant IL-17A (30 ng/ml) or IL-17F (30 ng/ml) for additional 48 h.

    Techniques: Activation Assay, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Expressing, Marker, Western Blot, Standard Deviation

    Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Journal: International Journal of Molecular Medicine

    Article Title: Adenovirus-mediated knockdown of activin A receptor type 2A attenuates immune-induced hepatic fibrosis in mice and inhibits interleukin-17-induced activation of primary hepatic stellate cells

    doi: 10.3892/ijmm.2018.3600

    Figure Lengend Snippet: Chronic treatment with Con A induces activation of activin and IL-17 signaling in mouse liver. (A) Experiment part 1: Mice were administered Con A (8 mg/kg/week) for up to 6 weeks, in order to generate a model of immune-associated liver fibrosis. (B–E) Serum and liver levels of activin A, IL-17A and IL-17F were detected in mice in the control and Con A groups using specific ELISA kits. (F) mRNA and (G) protein expression levels of ACVR2A were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Data are expressed as the means ± standard deviation. (H) Experiment part 2: Mice in the Con A groups were administered two injections of Ad-ACVR2A shRNA or Ad-NC shRNA at the indicated timepoints. (I–K) Protein expression levels of ACVR2A were analyzed in mouse liver tissues collected from the various groups. Data are expressed as the means ± standard error. ACVR2A, activin A receptor type 2A; Con A, concanavalin A; IL-17, interleukin-17; NC, negative control; shRNA, short hairpin RNA.

    Article Snippet: Recombinant murine IL-17A and IL-17F (PeproTech, Inc., Rocky Hill, NJ, USA) at 10, 30 or 100 ng/ml were used to stimulate mHSCs for 1, 3, 6, 12, 24 or 48 h. For some experiments, the mHSCs were infected with Ad-ACVR2A shRNA or Ad-NC shRNA (multiplicity of infection, both 50) for 24 h, and were then stimulated with recombinant IL-17A (30 ng/ml) or IL-17F (30 ng/ml) for additional 48 h.

    Techniques: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, shRNA, Negative Control

    Release of IL-18 in splenocytes from ICE KO mice. Splenocytes from WT or ICE KO mice were cultured for 24 hours in the absence of any exogenous stimulation. IL-18 levels were measured in supernatants. Data are mean ± SEM of 8 mice per group. ** P

    Journal: Journal of Clinical Investigation

    Article Title: IL-12-induced IFN-? is dependent on caspase-1 processing of the IL-18 precursor

    doi:

    Figure Lengend Snippet: Release of IL-18 in splenocytes from ICE KO mice. Splenocytes from WT or ICE KO mice were cultured for 24 hours in the absence of any exogenous stimulation. IL-18 levels were measured in supernatants. Data are mean ± SEM of 8 mice per group. ** P

    Article Snippet: Tubes were shaken for 15 minutes at room temperature, after which 25 μL of samples or standard concentrations of recombinant murine IL-18 (PeproTech Inc., Rocky Hill, New Jersey, USA) was added.

    Techniques: Mouse Assay, Cell Culture

    Constitutive expression of pro–IL-18 in the spleens and livers on untreated mice. Spleen and liver homogenates from 3 untreated mice were subjected to Western blot analysis with a specific rabbit anti-murine IL-18 antiserum. Recombinant murine pro–IL-18 and mature IL-18 were used as standards.

    Journal: Journal of Clinical Investigation

    Article Title: IL-12-induced IFN-? is dependent on caspase-1 processing of the IL-18 precursor

    doi:

    Figure Lengend Snippet: Constitutive expression of pro–IL-18 in the spleens and livers on untreated mice. Spleen and liver homogenates from 3 untreated mice were subjected to Western blot analysis with a specific rabbit anti-murine IL-18 antiserum. Recombinant murine pro–IL-18 and mature IL-18 were used as standards.

    Article Snippet: Tubes were shaken for 15 minutes at room temperature, after which 25 μL of samples or standard concentrations of recombinant murine IL-18 (PeproTech Inc., Rocky Hill, New Jersey, USA) was added.

    Techniques: Expressing, Mouse Assay, Western Blot, Recombinant

    Increase in serum IL-18 levels after IL-12 administration. WT and ICE KO mice received 4 daily intraperitoneal injections of IL-12 (100 or 400 ng/mouse) or vehicle. Two hours after the fourth injection, blood was collected and serum was prepared. Data are mean ± SEM of 10 mice per group. * P

    Journal: Journal of Clinical Investigation

    Article Title: IL-12-induced IFN-? is dependent on caspase-1 processing of the IL-18 precursor

    doi:

    Figure Lengend Snippet: Increase in serum IL-18 levels after IL-12 administration. WT and ICE KO mice received 4 daily intraperitoneal injections of IL-12 (100 or 400 ng/mouse) or vehicle. Two hours after the fourth injection, blood was collected and serum was prepared. Data are mean ± SEM of 10 mice per group. * P

    Article Snippet: Tubes were shaken for 15 minutes at room temperature, after which 25 μL of samples or standard concentrations of recombinant murine IL-18 (PeproTech Inc., Rocky Hill, New Jersey, USA) was added.

    Techniques: Mouse Assay, Injection

    Neutralization of IL-18 reduces IL-12–induced IFN-γ levels in vivo. Mice received 4 daily intraperitoneal injections of IL-12 (100 ng/mouse). One hour before the first and the third injection, the antibody-treated group received an intraperitoneal injection of 200 μL of rabbit anti–IL-18 antiserum, whereas the control group received the same amount of NRS. Two hours after the fourth injection, blood was collected and serum was prepared for measurement of IFN-γ. Data are mean ± SEM of 5 mice per group. ** P

    Journal: Journal of Clinical Investigation

    Article Title: IL-12-induced IFN-? is dependent on caspase-1 processing of the IL-18 precursor

    doi:

    Figure Lengend Snippet: Neutralization of IL-18 reduces IL-12–induced IFN-γ levels in vivo. Mice received 4 daily intraperitoneal injections of IL-12 (100 ng/mouse). One hour before the first and the third injection, the antibody-treated group received an intraperitoneal injection of 200 μL of rabbit anti–IL-18 antiserum, whereas the control group received the same amount of NRS. Two hours after the fourth injection, blood was collected and serum was prepared for measurement of IFN-γ. Data are mean ± SEM of 5 mice per group. ** P

    Article Snippet: Tubes were shaken for 15 minutes at room temperature, after which 25 μL of samples or standard concentrations of recombinant murine IL-18 (PeproTech Inc., Rocky Hill, New Jersey, USA) was added.

    Techniques: Neutralization, In Vivo, Mouse Assay, Injection

    IL-18 production in unstimulated and IL-12–stimulated splenocytes. Splenocytes were cultured in RPMI-FBS for increasing amounts of time without added exogenous stimulation, or with IL-12 at 10 ng/mL. Cell-associated and released IL-18 levels were measured. Data are mean ± SEM of 9 mice per group.

    Journal: Journal of Clinical Investigation

    Article Title: IL-12-induced IFN-? is dependent on caspase-1 processing of the IL-18 precursor

    doi:

    Figure Lengend Snippet: IL-18 production in unstimulated and IL-12–stimulated splenocytes. Splenocytes were cultured in RPMI-FBS for increasing amounts of time without added exogenous stimulation, or with IL-12 at 10 ng/mL. Cell-associated and released IL-18 levels were measured. Data are mean ± SEM of 9 mice per group.

    Article Snippet: Tubes were shaken for 15 minutes at room temperature, after which 25 μL of samples or standard concentrations of recombinant murine IL-18 (PeproTech Inc., Rocky Hill, New Jersey, USA) was added.

    Techniques: Cell Culture, Mouse Assay

    Effect of IL-18 neutralization on the induction of IFN-γ by IL-12 in vitro. Splenocytes were cultured for 48 hours with increasing concentrations of IL-12 (0.1–10 ng/mL) in the presence or absence of either neutralizing anti–IL-18 IgG or control IgG (50 μg/mL). IFN-γ levels were measured in the supernatants. Data are mean ± SEM of 3 mice per group and are representative of 1 experiment out of 3 performed. * P

    Journal: Journal of Clinical Investigation

    Article Title: IL-12-induced IFN-? is dependent on caspase-1 processing of the IL-18 precursor

    doi:

    Figure Lengend Snippet: Effect of IL-18 neutralization on the induction of IFN-γ by IL-12 in vitro. Splenocytes were cultured for 48 hours with increasing concentrations of IL-12 (0.1–10 ng/mL) in the presence or absence of either neutralizing anti–IL-18 IgG or control IgG (50 μg/mL). IFN-γ levels were measured in the supernatants. Data are mean ± SEM of 3 mice per group and are representative of 1 experiment out of 3 performed. * P

    Article Snippet: Tubes were shaken for 15 minutes at room temperature, after which 25 μL of samples or standard concentrations of recombinant murine IL-18 (PeproTech Inc., Rocky Hill, New Jersey, USA) was added.

    Techniques: Neutralization, In Vitro, Cell Culture, Mouse Assay

    Short hairpin RNA design, interference efficiency assay, and lentivirus transduction. A: DNA sequence analysis of pGCSIL/Interleukin-17A receptor (IL-17RA) short hairpin RNA (shRNA) 1; B: Lentiviral-mediated IL-17RA shRNA 1, 2, 3, and 4 (shRNA 1, 2, 3, and 4) inhibited IL-17RA mRNA expression in hepatic stellate cells (HSCs). HSCs infected with lentivirus at a multiplicity of infection of 10 for 72 h, and IL-17RA mRNA expression was quantified by polymerase chain reaction. IL-17RA shRNA 1 was the most efficient silencing tool for IL-17RA. b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Lentiviral vector-mediated down-regulation of IL-17A receptor in hepatic stellate cells results in decreased secretion of IL-6

    doi: 10.3748/wjg.v18.i28.3696

    Figure Lengend Snippet: Short hairpin RNA design, interference efficiency assay, and lentivirus transduction. A: DNA sequence analysis of pGCSIL/Interleukin-17A receptor (IL-17RA) short hairpin RNA (shRNA) 1; B: Lentiviral-mediated IL-17RA shRNA 1, 2, 3, and 4 (shRNA 1, 2, 3, and 4) inhibited IL-17RA mRNA expression in hepatic stellate cells (HSCs). HSCs infected with lentivirus at a multiplicity of infection of 10 for 72 h, and IL-17RA mRNA expression was quantified by polymerase chain reaction. IL-17RA shRNA 1 was the most efficient silencing tool for IL-17RA. b P

    Article Snippet: Recombinant murine IL-17A was purchased from Peprothech (Princeton Business Park, NJ).

    Techniques: shRNA, Transduction, Sequencing, Expressing, Infection, Polymerase Chain Reaction

    Mitogen activated protein kinases pathway involved in interleukin 17A induced interleukin 6 expression. A: Secretion of interleukin (IL)-6 in hepatic stellate cells (HSCs) induced by interleukin 17A (IL-17A) was determined using enzyme-linked immunosorbent assay. b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Lentiviral vector-mediated down-regulation of IL-17A receptor in hepatic stellate cells results in decreased secretion of IL-6

    doi: 10.3748/wjg.v18.i28.3696

    Figure Lengend Snippet: Mitogen activated protein kinases pathway involved in interleukin 17A induced interleukin 6 expression. A: Secretion of interleukin (IL)-6 in hepatic stellate cells (HSCs) induced by interleukin 17A (IL-17A) was determined using enzyme-linked immunosorbent assay. b P

    Article Snippet: Recombinant murine IL-17A was purchased from Peprothech (Princeton Business Park, NJ).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Lentiviral-mediated interleukin 17A receptor short hairpin RNA arrested interleukin 6 expression, partly through suppressing phosphorylation of p38 mitogen activated protein kinases and extracellular regulated protein kinases 1/2. A: Lentiviral-mediated interleukin (IL)-17A receptor short hairpin RNA (shRNA) 1 [at multiplicity of infection (MOI) = 10 for 72 h] inhibited the expression of IL-17RA in hepatic stellate cells (HSCs); B: Lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression in HSCs. HSCs were infected with IL-17RA shRNA 1 or random shRNA lentivirus at MOI = 10 for 72 h, and followed by 3 h exposure to IL-17A (100 ng/mL). b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Lentiviral vector-mediated down-regulation of IL-17A receptor in hepatic stellate cells results in decreased secretion of IL-6

    doi: 10.3748/wjg.v18.i28.3696

    Figure Lengend Snippet: Lentiviral-mediated interleukin 17A receptor short hairpin RNA arrested interleukin 6 expression, partly through suppressing phosphorylation of p38 mitogen activated protein kinases and extracellular regulated protein kinases 1/2. A: Lentiviral-mediated interleukin (IL)-17A receptor short hairpin RNA (shRNA) 1 [at multiplicity of infection (MOI) = 10 for 72 h] inhibited the expression of IL-17RA in hepatic stellate cells (HSCs); B: Lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression in HSCs. HSCs were infected with IL-17RA shRNA 1 or random shRNA lentivirus at MOI = 10 for 72 h, and followed by 3 h exposure to IL-17A (100 ng/mL). b P

    Article Snippet: Recombinant murine IL-17A was purchased from Peprothech (Princeton Business Park, NJ).

    Techniques: shRNA, Expressing, Infection