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recombinant mouse human il-36γ  (R&D Systems)


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    R&D Systems recombinant mouse human il-36γ
    Effects of <t>IL-36γ</t> on anchorage-independent growth and epithelial cell transformation in vitro. ( a ) JB6 Cl41 cells were treated with the indicated concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( b , c ) JB6 Cl41 cells were treated with different concentrations of IL-36γ as indicated and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( b ), followed by a calculation of the average colony numbers and sizes (diameter > 80 μm) ( c ). ( d ) MCF7 cells were treated with various concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( e , f ) MCF7 cells were treated with the indicated concentrations of IL-36γ and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( e ), followed by a calculation of the average colony numbers and sizes (diameter > 100 μm) ( f ). Error bars represent the mean ± standard deviation (S.D.) of triplicate measurements from at least three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control groups). In b and e, magnification, 100×.
    Recombinant Mouse Human Il 36γ, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+mouse+human+il-36%CE%B3/il+36%CE%B3/pmc09367291-46-3-8
    Average 90 stars, based on 1 article reviews
    recombinant mouse human il-36γ - by Bioz Stars, 2026-09
    90/100 stars

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    1) Product Images from "Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis"

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    Journal: Cancers

    doi: 10.3390/cancers14153654

    Effects of IL-36γ on anchorage-independent growth and epithelial cell transformation in vitro. ( a ) JB6 Cl41 cells were treated with the indicated concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( b , c ) JB6 Cl41 cells were treated with different concentrations of IL-36γ as indicated and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( b ), followed by a calculation of the average colony numbers and sizes (diameter > 80 μm) ( c ). ( d ) MCF7 cells were treated with various concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( e , f ) MCF7 cells were treated with the indicated concentrations of IL-36γ and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( e ), followed by a calculation of the average colony numbers and sizes (diameter > 100 μm) ( f ). Error bars represent the mean ± standard deviation (S.D.) of triplicate measurements from at least three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control groups). In b and e, magnification, 100×.
    Figure Legend Snippet: Effects of IL-36γ on anchorage-independent growth and epithelial cell transformation in vitro. ( a ) JB6 Cl41 cells were treated with the indicated concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( b , c ) JB6 Cl41 cells were treated with different concentrations of IL-36γ as indicated and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( b ), followed by a calculation of the average colony numbers and sizes (diameter > 80 μm) ( c ). ( d ) MCF7 cells were treated with various concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( e , f ) MCF7 cells were treated with the indicated concentrations of IL-36γ and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( e ), followed by a calculation of the average colony numbers and sizes (diameter > 100 μm) ( f ). Error bars represent the mean ± standard deviation (S.D.) of triplicate measurements from at least three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control groups). In b and e, magnification, 100×.

    Techniques Used: Transformation Assay, In Vitro, BrdU Incorporation Assay, Incubation, Standard Deviation

    Effects of IL-36γ on the MEK/ERK and JNK/c-Jun signaling pathway in JB6 Cl41 cells. ( a – d ) Cells were treated with the indicated concentrations of IL-36γ for 30 min ( a , c ) or 10 ng/mL IL-36γ for the indicated times ( b , d ). Proteins in the whole-cell lysates were analyzed by Western blotting. ( e ) Cells were transfected with mouse sgCtrl and sgIL-36R. After 48 h of transfection, cells were serum-starved for 24 h prior to treatment with 10 ng/mL of IL-36γ. After 15 min, total protein was collected, and the levels of the indicated protein were determined by Western blotting using specific antibodies. ( f , g ) Cells were pre-treated with the indicated concentrations of PD98059 ( f ) and SP600125 ( g ) for 12 h and then treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were analyzed by Western blotting. ( a – g ) Representative blots are shown from at least three independent experiments. 3.3. PIN1 Enhances AP-1 Activity and IL-36γ-Induced Transformation of JB6 Cl41 Cells. The full uncropped blots for can be found in .
    Figure Legend Snippet: Effects of IL-36γ on the MEK/ERK and JNK/c-Jun signaling pathway in JB6 Cl41 cells. ( a – d ) Cells were treated with the indicated concentrations of IL-36γ for 30 min ( a , c ) or 10 ng/mL IL-36γ for the indicated times ( b , d ). Proteins in the whole-cell lysates were analyzed by Western blotting. ( e ) Cells were transfected with mouse sgCtrl and sgIL-36R. After 48 h of transfection, cells were serum-starved for 24 h prior to treatment with 10 ng/mL of IL-36γ. After 15 min, total protein was collected, and the levels of the indicated protein were determined by Western blotting using specific antibodies. ( f , g ) Cells were pre-treated with the indicated concentrations of PD98059 ( f ) and SP600125 ( g ) for 12 h and then treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were analyzed by Western blotting. ( a – g ) Representative blots are shown from at least three independent experiments. 3.3. PIN1 Enhances AP-1 Activity and IL-36γ-Induced Transformation of JB6 Cl41 Cells. The full uncropped blots for can be found in .

    Techniques Used: Western Blot, Transfection, Activity Assay, Transformation Assay

    Effects of juglone in IL-36γ-induced AP-1 activity and cellular transformation of JB6 Cl41 cells. ( a , b ) Cells were transfected with mock and XP-PIN1 ( a ) or mouse sgCtrl and sgPIN1 ( b ), respectively. After 48 h of transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were determined by Western blotting. ( c ) Cells were exposed to the indicated amount of juglone for 12 h, prior to 15 min of treatment with 10 ng/mL IL-36γ. The levels of the indicated proteins were determined by Western blotting using specific antibodies. ( d – f ) The luciferase reporters c-Fos-Luc ( d ), c-Jun-Luc ( e ), and AP-1-Luc ( f ) were co-transfected along with the pRL-TK vector into cells. Cells were serum-starved for 24 h, following 24 h of transfection prior to dose-dependent treatment with IL-36γ for 24 h. ( g ) The luciferase reporter AP-1-Luc was co-transfected with pRL-TK vector into cells. Cells were pre-treated with the indicated doses of juglone, followed by 24 h of serum starvation and then exposed to 10 ng/mL IL-36γ for 24 h. ( h , i ) Cells were treated with 10 ng/mL IL-36γ either alone or in combination with 50 μM juglone in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative pictures of the colonies from three separate experiments ( h ) and the quantification of the colony sizes and numbers ( i ). Magnification, 100×. Error bars indicate the mean ± S.D. of triplicate measurements. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group). The full uncropped blots for can be found in .
    Figure Legend Snippet: Effects of juglone in IL-36γ-induced AP-1 activity and cellular transformation of JB6 Cl41 cells. ( a , b ) Cells were transfected with mock and XP-PIN1 ( a ) or mouse sgCtrl and sgPIN1 ( b ), respectively. After 48 h of transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were determined by Western blotting. ( c ) Cells were exposed to the indicated amount of juglone for 12 h, prior to 15 min of treatment with 10 ng/mL IL-36γ. The levels of the indicated proteins were determined by Western blotting using specific antibodies. ( d – f ) The luciferase reporters c-Fos-Luc ( d ), c-Jun-Luc ( e ), and AP-1-Luc ( f ) were co-transfected along with the pRL-TK vector into cells. Cells were serum-starved for 24 h, following 24 h of transfection prior to dose-dependent treatment with IL-36γ for 24 h. ( g ) The luciferase reporter AP-1-Luc was co-transfected with pRL-TK vector into cells. Cells were pre-treated with the indicated doses of juglone, followed by 24 h of serum starvation and then exposed to 10 ng/mL IL-36γ for 24 h. ( h , i ) Cells were treated with 10 ng/mL IL-36γ either alone or in combination with 50 μM juglone in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative pictures of the colonies from three separate experiments ( h ) and the quantification of the colony sizes and numbers ( i ). Magnification, 100×. Error bars indicate the mean ± S.D. of triplicate measurements. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group). The full uncropped blots for can be found in .

    Techniques Used: Activity Assay, Transformation Assay, Transfection, Western Blot, Luciferase, Plasmid Preparation, Incubation

    Role of PIN1 in the IL-36γ-induced MEK/ERK and JNK/c-Jun signaling pathways in MCF7 cells. ( a , b ) Cells were treated with the indicated concentration of IL-36γ for 30 min ( a ) or treated with 10 ng/mL IL-36γ for the indicated time ( b ). ( c ) Cells were transfected with human sgCtrl and sgIL-36R. After 48 h, cells were serum-starved for 24 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysate were determined by Western blotting. ( d , e ) Cells were transfected with mock and XP-PIN1 ( d ) or sgCtrl and sgPIN1 ( e ), respectively. At 48 h after transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. ( f ) Cells were pre-treated with the indicated doses of juglone for 12 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysates were analyzed by Western blotting. The blots shown are representative of at least three independent experiments.The full uncropped blots for can be found in .
    Figure Legend Snippet: Role of PIN1 in the IL-36γ-induced MEK/ERK and JNK/c-Jun signaling pathways in MCF7 cells. ( a , b ) Cells were treated with the indicated concentration of IL-36γ for 30 min ( a ) or treated with 10 ng/mL IL-36γ for the indicated time ( b ). ( c ) Cells were transfected with human sgCtrl and sgIL-36R. After 48 h, cells were serum-starved for 24 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysate were determined by Western blotting. ( d , e ) Cells were transfected with mock and XP-PIN1 ( d ) or sgCtrl and sgPIN1 ( e ), respectively. At 48 h after transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. ( f ) Cells were pre-treated with the indicated doses of juglone for 12 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysates were analyzed by Western blotting. The blots shown are representative of at least three independent experiments.The full uncropped blots for can be found in .

    Techniques Used: Concentration Assay, Transfection, Western Blot

    Role of PIN1 in IL-36γ-induced AP-1 activity and epithelial cell transformation. ( a – c ) The luciferase reporters c-Fos-Luc ( a ), c-Jun-Luc ( b ), and AP-1-Luc ( c ) were co-transfected with the pRL-TK vector into cells. Following 24 h of transfection, cells were serum-starved for 24 h prior to dose-dependent treatment with IL-36γ for 24 h. ( d ) The luciferase reporter AP-1-Luc and sgCtrl or AP-1-Luc and sgPIN1 were co-transfected into cells and incubated for 24 h. After being serum-starved for 24 h, cells were exposed to 10 ng/mL IL-36γ for another 24 h. ( e ) sgCtrl and sgPIN1 were transfected into cells and incubated for 48 h. Then, cells were treated with or without 10 ng/mL IL-36γ in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Magnification, 100×. ( f ) Representative images from three separate experiments are presented (left), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm, right). Error bars indicate the mean ± S.D. of triplicate measurements from three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group).
    Figure Legend Snippet: Role of PIN1 in IL-36γ-induced AP-1 activity and epithelial cell transformation. ( a – c ) The luciferase reporters c-Fos-Luc ( a ), c-Jun-Luc ( b ), and AP-1-Luc ( c ) were co-transfected with the pRL-TK vector into cells. Following 24 h of transfection, cells were serum-starved for 24 h prior to dose-dependent treatment with IL-36γ for 24 h. ( d ) The luciferase reporter AP-1-Luc and sgCtrl or AP-1-Luc and sgPIN1 were co-transfected into cells and incubated for 24 h. After being serum-starved for 24 h, cells were exposed to 10 ng/mL IL-36γ for another 24 h. ( e ) sgCtrl and sgPIN1 were transfected into cells and incubated for 48 h. Then, cells were treated with or without 10 ng/mL IL-36γ in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Magnification, 100×. ( f ) Representative images from three separate experiments are presented (left), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm, right). Error bars indicate the mean ± S.D. of triplicate measurements from three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group).

    Techniques Used: Activity Assay, Transformation Assay, Luciferase, Transfection, Plasmid Preparation, Incubation

    Role of PIN1 in IL-36γ-induced cell proliferation and mammary gland tumorigenesis. ( a ) 4T1 cells were seeded and treated with IL-36γ as indicated. Cell proliferation was estimated using the BrdU incorporation assay. ( b , c ) 4T1 cells were treated with the indicated doses of IL-36γ, subjected to a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( b ), followed by a calculation of the average colony numbers and sizes (diameter >200 μm) ( c ). ( d , e ) 4T1 cells were treated with the indicated concentrations of IL-36γ in the presence or absence of juglone, grown in a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( d ), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm) ( e ). ( f , g ) WT 4T1 and PIN1 KO 4T1 cells were injected into the mammary gland of BALB/c mice in the presence or absence of 100 ng/mL IL-36γ and allowed to grow until tumors were formed. Shown are representative images of tumor ( f ), measured tumor weights, and tumor volumes ( g ). Error bars indicate the mean ± S.D. of triplicate measurements from three independent experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group, respectively). In b and d, magnification, 100×.
    Figure Legend Snippet: Role of PIN1 in IL-36γ-induced cell proliferation and mammary gland tumorigenesis. ( a ) 4T1 cells were seeded and treated with IL-36γ as indicated. Cell proliferation was estimated using the BrdU incorporation assay. ( b , c ) 4T1 cells were treated with the indicated doses of IL-36γ, subjected to a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( b ), followed by a calculation of the average colony numbers and sizes (diameter >200 μm) ( c ). ( d , e ) 4T1 cells were treated with the indicated concentrations of IL-36γ in the presence or absence of juglone, grown in a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( d ), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm) ( e ). ( f , g ) WT 4T1 and PIN1 KO 4T1 cells were injected into the mammary gland of BALB/c mice in the presence or absence of 100 ng/mL IL-36γ and allowed to grow until tumors were formed. Shown are representative images of tumor ( f ), measured tumor weights, and tumor volumes ( g ). Error bars indicate the mean ± S.D. of triplicate measurements from three independent experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group, respectively). In b and d, magnification, 100×.

    Techniques Used: BrdU Incorporation Assay, Incubation, Injection



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    recombinant mouse human il-36γ  (R&D Systems)
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    R&D Systems recombinant mouse human il-36γ
    Effects of <t>IL-36γ</t> on anchorage-independent growth and epithelial cell transformation in vitro. ( a ) JB6 Cl41 cells were treated with the indicated concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( b , c ) JB6 Cl41 cells were treated with different concentrations of IL-36γ as indicated and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( b ), followed by a calculation of the average colony numbers and sizes (diameter > 80 μm) ( c ). ( d ) MCF7 cells were treated with various concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( e , f ) MCF7 cells were treated with the indicated concentrations of IL-36γ and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( e ), followed by a calculation of the average colony numbers and sizes (diameter > 100 μm) ( f ). Error bars represent the mean ± standard deviation (S.D.) of triplicate measurements from at least three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control groups). In b and e, magnification, 100×.
    Recombinant Mouse Human Il 36γ, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+mouse+human+il-36%CE%B3/il+36%CE%B3/pmc09367291-46-3-8
    Average 90 stars, based on 1 article reviews
    recombinant mouse human il-36γ - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier

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    Effects of IL-36γ on anchorage-independent growth and epithelial cell transformation in vitro. ( a ) JB6 Cl41 cells were treated with the indicated concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( b , c ) JB6 Cl41 cells were treated with different concentrations of IL-36γ as indicated and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( b ), followed by a calculation of the average colony numbers and sizes (diameter > 80 μm) ( c ). ( d ) MCF7 cells were treated with various concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( e , f ) MCF7 cells were treated with the indicated concentrations of IL-36γ and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( e ), followed by a calculation of the average colony numbers and sizes (diameter > 100 μm) ( f ). Error bars represent the mean ± standard deviation (S.D.) of triplicate measurements from at least three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control groups). In b and e, magnification, 100×.

    Journal: Cancers

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    doi: 10.3390/cancers14153654

    Figure Lengend Snippet: Effects of IL-36γ on anchorage-independent growth and epithelial cell transformation in vitro. ( a ) JB6 Cl41 cells were treated with the indicated concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( b , c ) JB6 Cl41 cells were treated with different concentrations of IL-36γ as indicated and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( b ), followed by a calculation of the average colony numbers and sizes (diameter > 80 μm) ( c ). ( d ) MCF7 cells were treated with various concentrations of IL-36γ for 48 h, and cell proliferation was determined using the BrdU incorporation assay. ( e , f ) MCF7 cells were treated with the indicated concentrations of IL-36γ and grown in a soft agar matrix by incubation at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are shown ( e ), followed by a calculation of the average colony numbers and sizes (diameter > 100 μm) ( f ). Error bars represent the mean ± standard deviation (S.D.) of triplicate measurements from at least three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control groups). In b and e, magnification, 100×.

    Article Snippet: Recombinant mouse and human IL-36γ were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Transformation Assay, In Vitro, BrdU Incorporation Assay, Incubation, Standard Deviation

    Effects of IL-36γ on the MEK/ERK and JNK/c-Jun signaling pathway in JB6 Cl41 cells. ( a – d ) Cells were treated with the indicated concentrations of IL-36γ for 30 min ( a , c ) or 10 ng/mL IL-36γ for the indicated times ( b , d ). Proteins in the whole-cell lysates were analyzed by Western blotting. ( e ) Cells were transfected with mouse sgCtrl and sgIL-36R. After 48 h of transfection, cells were serum-starved for 24 h prior to treatment with 10 ng/mL of IL-36γ. After 15 min, total protein was collected, and the levels of the indicated protein were determined by Western blotting using specific antibodies. ( f , g ) Cells were pre-treated with the indicated concentrations of PD98059 ( f ) and SP600125 ( g ) for 12 h and then treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were analyzed by Western blotting. ( a – g ) Representative blots are shown from at least three independent experiments. 3.3. PIN1 Enhances AP-1 Activity and IL-36γ-Induced Transformation of JB6 Cl41 Cells. The full uncropped blots for can be found in .

    Journal: Cancers

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    doi: 10.3390/cancers14153654

    Figure Lengend Snippet: Effects of IL-36γ on the MEK/ERK and JNK/c-Jun signaling pathway in JB6 Cl41 cells. ( a – d ) Cells were treated with the indicated concentrations of IL-36γ for 30 min ( a , c ) or 10 ng/mL IL-36γ for the indicated times ( b , d ). Proteins in the whole-cell lysates were analyzed by Western blotting. ( e ) Cells were transfected with mouse sgCtrl and sgIL-36R. After 48 h of transfection, cells were serum-starved for 24 h prior to treatment with 10 ng/mL of IL-36γ. After 15 min, total protein was collected, and the levels of the indicated protein were determined by Western blotting using specific antibodies. ( f , g ) Cells were pre-treated with the indicated concentrations of PD98059 ( f ) and SP600125 ( g ) for 12 h and then treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were analyzed by Western blotting. ( a – g ) Representative blots are shown from at least three independent experiments. 3.3. PIN1 Enhances AP-1 Activity and IL-36γ-Induced Transformation of JB6 Cl41 Cells. The full uncropped blots for can be found in .

    Article Snippet: Recombinant mouse and human IL-36γ were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Western Blot, Transfection, Activity Assay, Transformation Assay

    Effects of juglone in IL-36γ-induced AP-1 activity and cellular transformation of JB6 Cl41 cells. ( a , b ) Cells were transfected with mock and XP-PIN1 ( a ) or mouse sgCtrl and sgPIN1 ( b ), respectively. After 48 h of transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were determined by Western blotting. ( c ) Cells were exposed to the indicated amount of juglone for 12 h, prior to 15 min of treatment with 10 ng/mL IL-36γ. The levels of the indicated proteins were determined by Western blotting using specific antibodies. ( d – f ) The luciferase reporters c-Fos-Luc ( d ), c-Jun-Luc ( e ), and AP-1-Luc ( f ) were co-transfected along with the pRL-TK vector into cells. Cells were serum-starved for 24 h, following 24 h of transfection prior to dose-dependent treatment with IL-36γ for 24 h. ( g ) The luciferase reporter AP-1-Luc was co-transfected with pRL-TK vector into cells. Cells were pre-treated with the indicated doses of juglone, followed by 24 h of serum starvation and then exposed to 10 ng/mL IL-36γ for 24 h. ( h , i ) Cells were treated with 10 ng/mL IL-36γ either alone or in combination with 50 μM juglone in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative pictures of the colonies from three separate experiments ( h ) and the quantification of the colony sizes and numbers ( i ). Magnification, 100×. Error bars indicate the mean ± S.D. of triplicate measurements. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group). The full uncropped blots for can be found in .

    Journal: Cancers

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    doi: 10.3390/cancers14153654

    Figure Lengend Snippet: Effects of juglone in IL-36γ-induced AP-1 activity and cellular transformation of JB6 Cl41 cells. ( a , b ) Cells were transfected with mock and XP-PIN1 ( a ) or mouse sgCtrl and sgPIN1 ( b ), respectively. After 48 h of transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. Proteins in the whole-cell lysates were determined by Western blotting. ( c ) Cells were exposed to the indicated amount of juglone for 12 h, prior to 15 min of treatment with 10 ng/mL IL-36γ. The levels of the indicated proteins were determined by Western blotting using specific antibodies. ( d – f ) The luciferase reporters c-Fos-Luc ( d ), c-Jun-Luc ( e ), and AP-1-Luc ( f ) were co-transfected along with the pRL-TK vector into cells. Cells were serum-starved for 24 h, following 24 h of transfection prior to dose-dependent treatment with IL-36γ for 24 h. ( g ) The luciferase reporter AP-1-Luc was co-transfected with pRL-TK vector into cells. Cells were pre-treated with the indicated doses of juglone, followed by 24 h of serum starvation and then exposed to 10 ng/mL IL-36γ for 24 h. ( h , i ) Cells were treated with 10 ng/mL IL-36γ either alone or in combination with 50 μM juglone in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative pictures of the colonies from three separate experiments ( h ) and the quantification of the colony sizes and numbers ( i ). Magnification, 100×. Error bars indicate the mean ± S.D. of triplicate measurements. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group). The full uncropped blots for can be found in .

    Article Snippet: Recombinant mouse and human IL-36γ were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Activity Assay, Transformation Assay, Transfection, Western Blot, Luciferase, Plasmid Preparation, Incubation

    Role of PIN1 in the IL-36γ-induced MEK/ERK and JNK/c-Jun signaling pathways in MCF7 cells. ( a , b ) Cells were treated with the indicated concentration of IL-36γ for 30 min ( a ) or treated with 10 ng/mL IL-36γ for the indicated time ( b ). ( c ) Cells were transfected with human sgCtrl and sgIL-36R. After 48 h, cells were serum-starved for 24 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysate were determined by Western blotting. ( d , e ) Cells were transfected with mock and XP-PIN1 ( d ) or sgCtrl and sgPIN1 ( e ), respectively. At 48 h after transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. ( f ) Cells were pre-treated with the indicated doses of juglone for 12 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysates were analyzed by Western blotting. The blots shown are representative of at least three independent experiments.The full uncropped blots for can be found in .

    Journal: Cancers

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    doi: 10.3390/cancers14153654

    Figure Lengend Snippet: Role of PIN1 in the IL-36γ-induced MEK/ERK and JNK/c-Jun signaling pathways in MCF7 cells. ( a , b ) Cells were treated with the indicated concentration of IL-36γ for 30 min ( a ) or treated with 10 ng/mL IL-36γ for the indicated time ( b ). ( c ) Cells were transfected with human sgCtrl and sgIL-36R. After 48 h, cells were serum-starved for 24 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysate were determined by Western blotting. ( d , e ) Cells were transfected with mock and XP-PIN1 ( d ) or sgCtrl and sgPIN1 ( e ), respectively. At 48 h after transfection, cells were serum-starved for 24 h and treated with 10 ng/mL IL-36γ for 15 min. ( f ) Cells were pre-treated with the indicated doses of juglone for 12 h prior to treatment with 10 ng/mL IL-36γ. After 15 min, cells were harvested and lysed. Proteins in the whole-cell lysates were analyzed by Western blotting. The blots shown are representative of at least three independent experiments.The full uncropped blots for can be found in .

    Article Snippet: Recombinant mouse and human IL-36γ were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Concentration Assay, Transfection, Western Blot

    Role of PIN1 in IL-36γ-induced AP-1 activity and epithelial cell transformation. ( a – c ) The luciferase reporters c-Fos-Luc ( a ), c-Jun-Luc ( b ), and AP-1-Luc ( c ) were co-transfected with the pRL-TK vector into cells. Following 24 h of transfection, cells were serum-starved for 24 h prior to dose-dependent treatment with IL-36γ for 24 h. ( d ) The luciferase reporter AP-1-Luc and sgCtrl or AP-1-Luc and sgPIN1 were co-transfected into cells and incubated for 24 h. After being serum-starved for 24 h, cells were exposed to 10 ng/mL IL-36γ for another 24 h. ( e ) sgCtrl and sgPIN1 were transfected into cells and incubated for 48 h. Then, cells were treated with or without 10 ng/mL IL-36γ in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Magnification, 100×. ( f ) Representative images from three separate experiments are presented (left), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm, right). Error bars indicate the mean ± S.D. of triplicate measurements from three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group).

    Journal: Cancers

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    doi: 10.3390/cancers14153654

    Figure Lengend Snippet: Role of PIN1 in IL-36γ-induced AP-1 activity and epithelial cell transformation. ( a – c ) The luciferase reporters c-Fos-Luc ( a ), c-Jun-Luc ( b ), and AP-1-Luc ( c ) were co-transfected with the pRL-TK vector into cells. Following 24 h of transfection, cells were serum-starved for 24 h prior to dose-dependent treatment with IL-36γ for 24 h. ( d ) The luciferase reporter AP-1-Luc and sgCtrl or AP-1-Luc and sgPIN1 were co-transfected into cells and incubated for 24 h. After being serum-starved for 24 h, cells were exposed to 10 ng/mL IL-36γ for another 24 h. ( e ) sgCtrl and sgPIN1 were transfected into cells and incubated for 48 h. Then, cells were treated with or without 10 ng/mL IL-36γ in a soft agar matrix and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Magnification, 100×. ( f ) Representative images from three separate experiments are presented (left), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm, right). Error bars indicate the mean ± S.D. of triplicate measurements from three individual experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group).

    Article Snippet: Recombinant mouse and human IL-36γ were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Activity Assay, Transformation Assay, Luciferase, Transfection, Plasmid Preparation, Incubation

    Role of PIN1 in IL-36γ-induced cell proliferation and mammary gland tumorigenesis. ( a ) 4T1 cells were seeded and treated with IL-36γ as indicated. Cell proliferation was estimated using the BrdU incorporation assay. ( b , c ) 4T1 cells were treated with the indicated doses of IL-36γ, subjected to a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( b ), followed by a calculation of the average colony numbers and sizes (diameter >200 μm) ( c ). ( d , e ) 4T1 cells were treated with the indicated concentrations of IL-36γ in the presence or absence of juglone, grown in a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( d ), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm) ( e ). ( f , g ) WT 4T1 and PIN1 KO 4T1 cells were injected into the mammary gland of BALB/c mice in the presence or absence of 100 ng/mL IL-36γ and allowed to grow until tumors were formed. Shown are representative images of tumor ( f ), measured tumor weights, and tumor volumes ( g ). Error bars indicate the mean ± S.D. of triplicate measurements from three independent experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group, respectively). In b and d, magnification, 100×.

    Journal: Cancers

    Article Title: Regulation of Interleukin-36γ/IL-36R Signaling Axis by PIN1 in Epithelial Cell Transformation and Breast Tumorigenesis

    doi: 10.3390/cancers14153654

    Figure Lengend Snippet: Role of PIN1 in IL-36γ-induced cell proliferation and mammary gland tumorigenesis. ( a ) 4T1 cells were seeded and treated with IL-36γ as indicated. Cell proliferation was estimated using the BrdU incorporation assay. ( b , c ) 4T1 cells were treated with the indicated doses of IL-36γ, subjected to a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( b ), followed by a calculation of the average colony numbers and sizes (diameter >200 μm) ( c ). ( d , e ) 4T1 cells were treated with the indicated concentrations of IL-36γ in the presence or absence of juglone, grown in a soft agar matrix, and incubated at 37 °C in a 5% CO 2 atmosphere for 14 days. Representative images from three separate experiments are presented ( d ), followed by a calculation of the average colony numbers and sizes (diameter > 200 μm) ( e ). ( f , g ) WT 4T1 and PIN1 KO 4T1 cells were injected into the mammary gland of BALB/c mice in the presence or absence of 100 ng/mL IL-36γ and allowed to grow until tumors were formed. Shown are representative images of tumor ( f ), measured tumor weights, and tumor volumes ( g ). Error bars indicate the mean ± S.D. of triplicate measurements from three independent experiments. Statistical analyses were performed using one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group or only-IL-36γ-treated group, respectively). In b and d, magnification, 100×.

    Article Snippet: Recombinant mouse and human IL-36γ were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: BrdU Incorporation Assay, Incubation, Injection