rabbit anti orexin a hcrt  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti orexin a hcrt
    Rabbit Anti Orexin A Hcrt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti orexin a hcrt/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    rabbit anti orexin a hcrt  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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    Cell Signaling Technology Inc rabbit anti orexin a hcrt
    Rabbit Anti Orexin A Hcrt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti orexin a hcrt/product/Cell Signaling Technology Inc
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    anti phospho β catenin ser33  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho β catenin ser33
    The effects of LGR5 overexpression on cell proliferation <t>and</t> <t>WNT/β-catenin</t> signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The optical density (OD) value was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).
    Anti Phospho β Catenin Ser33, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho β catenin ser33/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho β catenin ser33 - by Bioz Stars, 2023-01
    94/100 stars

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    1) Product Images from "LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling"

    Article Title: LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19041036

    The effects of LGR5 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The optical density (OD) value was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).
    Figure Legend Snippet: The effects of LGR5 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The optical density (OD) value was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Techniques Used: Over Expression, Expressing, Cell Counting, MTT Assay, Western Blot

    The effects of BMI1 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The OD value was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).
    Figure Legend Snippet: The effects of BMI1 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The OD value was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Techniques Used: Over Expression, Expressing, Cell Counting, MTT Assay, Western Blot

    The effects of rhWNT3A protein supplementation on cell proliferation and protein expression in IPEC-J2 cells. ( A ) The OD values were higher in the 1.5- and 3.0-nmol/L WNT3A groups than in the control group at 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( B ) The levels of WNT/β-catenin signaling-related proteins, LGR5 and BMI1 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).
    Figure Legend Snippet: The effects of rhWNT3A protein supplementation on cell proliferation and protein expression in IPEC-J2 cells. ( A ) The OD values were higher in the 1.5- and 3.0-nmol/L WNT3A groups than in the control group at 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( B ) The levels of WNT/β-catenin signaling-related proteins, LGR5 and BMI1 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Techniques Used: Expressing, MTT Assay, Western Blot

    The effects of XAV939 supplementation on cell proliferation and protein expression in LGR5 -overexpressing or BMI1 -overexpressing IPEC-J2 cells. ( A ) The OD values were lower in the 5-, 10- or 15-μmol/L XAV939 groups than in the BMI1 -overexpressing group at 24, 48 and 72 h after treatment, as assessed by the MTT assay ( n = 20); ( B ) The levels of AXIN2, β-catenin and BMI1 were assessed by Western blot ( n = 3); ( C ) The OD value was lower in the 10-μmol/L XAV939 group than in the LGR5 -overexpressing group at 24 and 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( D ) The levels of WNT/β-catenin signaling-related proteins and LGR5 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).
    Figure Legend Snippet: The effects of XAV939 supplementation on cell proliferation and protein expression in LGR5 -overexpressing or BMI1 -overexpressing IPEC-J2 cells. ( A ) The OD values were lower in the 5-, 10- or 15-μmol/L XAV939 groups than in the BMI1 -overexpressing group at 24, 48 and 72 h after treatment, as assessed by the MTT assay ( n = 20); ( B ) The levels of AXIN2, β-catenin and BMI1 were assessed by Western blot ( n = 3); ( C ) The OD value was lower in the 10-μmol/L XAV939 group than in the LGR5 -overexpressing group at 24 and 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( D ) The levels of WNT/β-catenin signaling-related proteins and LGR5 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Techniques Used: Expressing, MTT Assay, Western Blot

    p β catenin s33  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p β catenin s33
    Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, <t>GSK3β,</t> <t>β-catenin,</t> and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.
    P β Catenin S33, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p β catenin s33/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    p β catenin s33 - by Bioz Stars, 2023-01
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    1) Product Images from "An Isoxazole Chalcone Derivative Enhances Melanogenesis in B16 Melanoma Cells via the Akt/GSK3β/β-Catenin Signaling Pathways"

    Article Title: An Isoxazole Chalcone Derivative Enhances Melanogenesis in B16 Melanoma Cells via the Akt/GSK3β/β-Catenin Signaling Pathways

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules22122077

    Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, GSK3β, β-catenin, and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.
    Figure Legend Snippet: Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, GSK3β, β-catenin, and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.

    Techniques Used: Transduction, Western Blot, Expressing

    PMPP promotes β-catenin nuclear translocation. B16 cells were treated with PMPP of 50 μM for 12 h, and the expression levels of proteins, including β-catenin in the cytoplasm and nucleus, were detected by using a Western blot. Equal protein loading amounts were confirmed by HSP 70 expression in the cytoplasm and Histone H3.1 expression in the nucleus. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. NS: No statistical difference, ** p < 0.01 compared with untreated cells.
    Figure Legend Snippet: PMPP promotes β-catenin nuclear translocation. B16 cells were treated with PMPP of 50 μM for 12 h, and the expression levels of proteins, including β-catenin in the cytoplasm and nucleus, were detected by using a Western blot. Equal protein loading amounts were confirmed by HSP 70 expression in the cytoplasm and Histone H3.1 expression in the nucleus. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. NS: No statistical difference, ** p < 0.01 compared with untreated cells.

    Techniques Used: Translocation Assay, Expressing, Western Blot

    Effects of PMPP on β-catenin, Microphthalmia-associated transcription factor (MITF), tyrosinase (TYR) protein expression and TYR activity, and melanin content in B16 cells treated with GSK3β or Akt inhibitor. ( A ) The GSK3β inhibitor increased PMPP-induced β-catenin expression. β-catenin was detected in B16 cells pre-treated with or without BIO (5 μM) and incubated with PMPP (50 μM) or not. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 versus untreated cells, # p < 0.05 versus PMPP alone; ( B ) Effects of inhibitors on PMPP-induced MITF and TYR expression. The expressions of MITF and TYR were determined by Western blot in B16 cells treated with PMPP or not in the presence or absence of BIO (5 μM) or Akt inhibitor IV (1 μM) for 48 h. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01 versus MITF control, # p < 0.05, ## p < 0.01 versus TYR control. GSK3β inhibitor enhanced PMPP-induced TYR activity ( C ) and melanin content ( D ). The Akt inhibitor suppressed PMPP-induced TYR activity ( E ) and melanin content ( F ). B16 cells pre-incubated with inhibitors (BIO 5 μM, AKT inhibitor IV 1 μM) were further incubated with 50 μM PMPP, then followed by an additional incubation for 24 h (tyrosinase activity) or 48 h (melanin content). Each percentage value in the treated cells was calculated with respect to that in the untreated cells. Values are expressed as the mean ± SD of three separate experiments. ** p < 0.01, *** p < 0.001 compared with control; # p < 0.05, ### p < 0.001 compared with PMPP stimulation.
    Figure Legend Snippet: Effects of PMPP on β-catenin, Microphthalmia-associated transcription factor (MITF), tyrosinase (TYR) protein expression and TYR activity, and melanin content in B16 cells treated with GSK3β or Akt inhibitor. ( A ) The GSK3β inhibitor increased PMPP-induced β-catenin expression. β-catenin was detected in B16 cells pre-treated with or without BIO (5 μM) and incubated with PMPP (50 μM) or not. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 versus untreated cells, # p < 0.05 versus PMPP alone; ( B ) Effects of inhibitors on PMPP-induced MITF and TYR expression. The expressions of MITF and TYR were determined by Western blot in B16 cells treated with PMPP or not in the presence or absence of BIO (5 μM) or Akt inhibitor IV (1 μM) for 48 h. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01 versus MITF control, # p < 0.05, ## p < 0.01 versus TYR control. GSK3β inhibitor enhanced PMPP-induced TYR activity ( C ) and melanin content ( D ). The Akt inhibitor suppressed PMPP-induced TYR activity ( E ) and melanin content ( F ). B16 cells pre-incubated with inhibitors (BIO 5 μM, AKT inhibitor IV 1 μM) were further incubated with 50 μM PMPP, then followed by an additional incubation for 24 h (tyrosinase activity) or 48 h (melanin content). Each percentage value in the treated cells was calculated with respect to that in the untreated cells. Values are expressed as the mean ± SD of three separate experiments. ** p < 0.01, *** p < 0.001 compared with control; # p < 0.05, ### p < 0.001 compared with PMPP stimulation.

    Techniques Used: Expressing, Activity Assay, Incubation, Western Blot

    The possible mechanism of PMPP activating melanin content in B16 cells. PMPP increases the phosphorylation of Akt, and thus probably increases the non-activated GSK3β, which inhibits the phosphorylation of β-catenin and ceases its degradation via proteasomes. β-catenin accumulating in cytoplasm up-regulates MITF content through promoting transcription with LEF/TCF (Lymphoidenhancer factor/T cell factor) after coming into nucleus. The augmentation of β-catenin probably leads to the increases in the transcription of MITF and the TYR family and the subsequent induction of melanin synthesis.
    Figure Legend Snippet: The possible mechanism of PMPP activating melanin content in B16 cells. PMPP increases the phosphorylation of Akt, and thus probably increases the non-activated GSK3β, which inhibits the phosphorylation of β-catenin and ceases its degradation via proteasomes. β-catenin accumulating in cytoplasm up-regulates MITF content through promoting transcription with LEF/TCF (Lymphoidenhancer factor/T cell factor) after coming into nucleus. The augmentation of β-catenin probably leads to the increases in the transcription of MITF and the TYR family and the subsequent induction of melanin synthesis.

    Techniques Used:

    anti p β catenin  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p β catenin
    Anti P β Catenin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti orexin a hcrt
    Rabbit Anti Orexin A Hcrt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho β catenin ser33
    The effects of LGR5 overexpression on cell proliferation <t>and</t> <t>WNT/β-catenin</t> signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The optical density (OD) value was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).
    Anti Phospho β Catenin Ser33, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc p β catenin s33
    Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, <t>GSK3β,</t> <t>β-catenin,</t> and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.
    P β Catenin S33, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p β catenin s33/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    Cell Signaling Technology Inc anti p β catenin
    Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, <t>GSK3β,</t> <t>β-catenin,</t> and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.
    Anti P β Catenin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p β catenin/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    anti p β catenin - by Bioz Stars, 2023-01
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    The effects of LGR5 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The optical density (OD) value was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Journal: International Journal of Molecular Sciences

    Article Title: LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

    doi: 10.3390/ijms19041036

    Figure Lengend Snippet: The effects of LGR5 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The optical density (OD) value was higher in the LGR5 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Article Snippet: The antibodies used in this study were as follows: anti-C-MYC (#5605) and anti-β-actin (#4970) from Cell Signaling Technology (Beverly, MA, USA); anti-phospho-β-catenin (Ser33) (sc-16743), anti-TCF4 (sc-13027), anti-GSK3β (sc-9166), anti-cyclin D1 (sc-753) and anti-BMI1 (sc-10745) from Santa Cruz (Dallas, TX, USA); anti-β-catenin (ab6302) and anti-AXIN2 (ab109307) from Abcam (Cambridge, MA, USA); anti-LGR5 from ABGENT (San Diego, CA, USA); and anti-rabbit IgG and anti-mouse IgG from Beijing Biosynthesis Biotechnology (Beijing, China).

    Techniques: Over Expression, Expressing, Cell Counting, MTT Assay, Western Blot

    The effects of BMI1 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The OD value was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Journal: International Journal of Molecular Sciences

    Article Title: LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

    doi: 10.3390/ijms19041036

    Figure Lengend Snippet: The effects of BMI1 overexpression on cell proliferation and WNT/β-catenin signaling-related protein expression in IPEC-J2 cells. ( A ) The cell number was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the cell count assay ( n = 3); ( B ) The OD value was higher in the BMI1 -overexpressing group than in the control group at 48, 72 and 96 h after seeding, as assessed by the MTT assay ( n = 20); and ( C ) The levels of WNT/β-catenin signaling-related proteins were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Article Snippet: The antibodies used in this study were as follows: anti-C-MYC (#5605) and anti-β-actin (#4970) from Cell Signaling Technology (Beverly, MA, USA); anti-phospho-β-catenin (Ser33) (sc-16743), anti-TCF4 (sc-13027), anti-GSK3β (sc-9166), anti-cyclin D1 (sc-753) and anti-BMI1 (sc-10745) from Santa Cruz (Dallas, TX, USA); anti-β-catenin (ab6302) and anti-AXIN2 (ab109307) from Abcam (Cambridge, MA, USA); anti-LGR5 from ABGENT (San Diego, CA, USA); and anti-rabbit IgG and anti-mouse IgG from Beijing Biosynthesis Biotechnology (Beijing, China).

    Techniques: Over Expression, Expressing, Cell Counting, MTT Assay, Western Blot

    The effects of rhWNT3A protein supplementation on cell proliferation and protein expression in IPEC-J2 cells. ( A ) The OD values were higher in the 1.5- and 3.0-nmol/L WNT3A groups than in the control group at 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( B ) The levels of WNT/β-catenin signaling-related proteins, LGR5 and BMI1 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Journal: International Journal of Molecular Sciences

    Article Title: LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

    doi: 10.3390/ijms19041036

    Figure Lengend Snippet: The effects of rhWNT3A protein supplementation on cell proliferation and protein expression in IPEC-J2 cells. ( A ) The OD values were higher in the 1.5- and 3.0-nmol/L WNT3A groups than in the control group at 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( B ) The levels of WNT/β-catenin signaling-related proteins, LGR5 and BMI1 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Article Snippet: The antibodies used in this study were as follows: anti-C-MYC (#5605) and anti-β-actin (#4970) from Cell Signaling Technology (Beverly, MA, USA); anti-phospho-β-catenin (Ser33) (sc-16743), anti-TCF4 (sc-13027), anti-GSK3β (sc-9166), anti-cyclin D1 (sc-753) and anti-BMI1 (sc-10745) from Santa Cruz (Dallas, TX, USA); anti-β-catenin (ab6302) and anti-AXIN2 (ab109307) from Abcam (Cambridge, MA, USA); anti-LGR5 from ABGENT (San Diego, CA, USA); and anti-rabbit IgG and anti-mouse IgG from Beijing Biosynthesis Biotechnology (Beijing, China).

    Techniques: Expressing, MTT Assay, Western Blot

    The effects of XAV939 supplementation on cell proliferation and protein expression in LGR5 -overexpressing or BMI1 -overexpressing IPEC-J2 cells. ( A ) The OD values were lower in the 5-, 10- or 15-μmol/L XAV939 groups than in the BMI1 -overexpressing group at 24, 48 and 72 h after treatment, as assessed by the MTT assay ( n = 20); ( B ) The levels of AXIN2, β-catenin and BMI1 were assessed by Western blot ( n = 3); ( C ) The OD value was lower in the 10-μmol/L XAV939 group than in the LGR5 -overexpressing group at 24 and 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( D ) The levels of WNT/β-catenin signaling-related proteins and LGR5 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Journal: International Journal of Molecular Sciences

    Article Title: LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

    doi: 10.3390/ijms19041036

    Figure Lengend Snippet: The effects of XAV939 supplementation on cell proliferation and protein expression in LGR5 -overexpressing or BMI1 -overexpressing IPEC-J2 cells. ( A ) The OD values were lower in the 5-, 10- or 15-μmol/L XAV939 groups than in the BMI1 -overexpressing group at 24, 48 and 72 h after treatment, as assessed by the MTT assay ( n = 20); ( B ) The levels of AXIN2, β-catenin and BMI1 were assessed by Western blot ( n = 3); ( C ) The OD value was lower in the 10-μmol/L XAV939 group than in the LGR5 -overexpressing group at 24 and 48 h after treatment, as assessed by the MTT assay ( n = 20); and ( D ) The levels of WNT/β-catenin signaling-related proteins and LGR5 were assessed by Western blot ( n = 3). The results were confirmed by three independent experiments per treatment. Representative results of the three independent experiments are shown. The bars are the means ± SE, * indicates a significant difference ( p < 0.05).

    Article Snippet: The antibodies used in this study were as follows: anti-C-MYC (#5605) and anti-β-actin (#4970) from Cell Signaling Technology (Beverly, MA, USA); anti-phospho-β-catenin (Ser33) (sc-16743), anti-TCF4 (sc-13027), anti-GSK3β (sc-9166), anti-cyclin D1 (sc-753) and anti-BMI1 (sc-10745) from Santa Cruz (Dallas, TX, USA); anti-β-catenin (ab6302) and anti-AXIN2 (ab109307) from Abcam (Cambridge, MA, USA); anti-LGR5 from ABGENT (San Diego, CA, USA); and anti-rabbit IgG and anti-mouse IgG from Beijing Biosynthesis Biotechnology (Beijing, China).

    Techniques: Expressing, MTT Assay, Western Blot

    Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, GSK3β, β-catenin, and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: An Isoxazole Chalcone Derivative Enhances Melanogenesis in B16 Melanoma Cells via the Akt/GSK3β/β-Catenin Signaling Pathways

    doi: 10.3390/molecules22122077

    Figure Lengend Snippet: Effects of PMPP on signal transduction proteins that participate in melanogenesis. B16 cells were treated with PMPP at 0, 2, 10, and 50 μM for 48 h, and the phosphorylation and total of Akt, GSK3β, β-catenin, and MITF were measured by Western blotting. Equal protein loading amounts were confirmed by β-actin expression.

    Article Snippet: AKT (#5373), p-AKT s473 (#9271), GSK3β (#9832), p-GSK3β s9 (#9323), β-catenin (#8480), and β-actin (#3700) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). p-β-catenin s33 (sc-16743-R), antibodies against TYR (sc-7833), TRP-1 (sc-25543), and TRP-2 (sc-25544) were bought from Santa Cruz Technology (Dallas, TX, USA).

    Techniques: Transduction, Western Blot, Expressing

    PMPP promotes β-catenin nuclear translocation. B16 cells were treated with PMPP of 50 μM for 12 h, and the expression levels of proteins, including β-catenin in the cytoplasm and nucleus, were detected by using a Western blot. Equal protein loading amounts were confirmed by HSP 70 expression in the cytoplasm and Histone H3.1 expression in the nucleus. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. NS: No statistical difference, ** p < 0.01 compared with untreated cells.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: An Isoxazole Chalcone Derivative Enhances Melanogenesis in B16 Melanoma Cells via the Akt/GSK3β/β-Catenin Signaling Pathways

    doi: 10.3390/molecules22122077

    Figure Lengend Snippet: PMPP promotes β-catenin nuclear translocation. B16 cells were treated with PMPP of 50 μM for 12 h, and the expression levels of proteins, including β-catenin in the cytoplasm and nucleus, were detected by using a Western blot. Equal protein loading amounts were confirmed by HSP 70 expression in the cytoplasm and Histone H3.1 expression in the nucleus. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. NS: No statistical difference, ** p < 0.01 compared with untreated cells.

    Article Snippet: AKT (#5373), p-AKT s473 (#9271), GSK3β (#9832), p-GSK3β s9 (#9323), β-catenin (#8480), and β-actin (#3700) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). p-β-catenin s33 (sc-16743-R), antibodies against TYR (sc-7833), TRP-1 (sc-25543), and TRP-2 (sc-25544) were bought from Santa Cruz Technology (Dallas, TX, USA).

    Techniques: Translocation Assay, Expressing, Western Blot

    Effects of PMPP on β-catenin, Microphthalmia-associated transcription factor (MITF), tyrosinase (TYR) protein expression and TYR activity, and melanin content in B16 cells treated with GSK3β or Akt inhibitor. ( A ) The GSK3β inhibitor increased PMPP-induced β-catenin expression. β-catenin was detected in B16 cells pre-treated with or without BIO (5 μM) and incubated with PMPP (50 μM) or not. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 versus untreated cells, # p < 0.05 versus PMPP alone; ( B ) Effects of inhibitors on PMPP-induced MITF and TYR expression. The expressions of MITF and TYR were determined by Western blot in B16 cells treated with PMPP or not in the presence or absence of BIO (5 μM) or Akt inhibitor IV (1 μM) for 48 h. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01 versus MITF control, # p < 0.05, ## p < 0.01 versus TYR control. GSK3β inhibitor enhanced PMPP-induced TYR activity ( C ) and melanin content ( D ). The Akt inhibitor suppressed PMPP-induced TYR activity ( E ) and melanin content ( F ). B16 cells pre-incubated with inhibitors (BIO 5 μM, AKT inhibitor IV 1 μM) were further incubated with 50 μM PMPP, then followed by an additional incubation for 24 h (tyrosinase activity) or 48 h (melanin content). Each percentage value in the treated cells was calculated with respect to that in the untreated cells. Values are expressed as the mean ± SD of three separate experiments. ** p < 0.01, *** p < 0.001 compared with control; # p < 0.05, ### p < 0.001 compared with PMPP stimulation.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: An Isoxazole Chalcone Derivative Enhances Melanogenesis in B16 Melanoma Cells via the Akt/GSK3β/β-Catenin Signaling Pathways

    doi: 10.3390/molecules22122077

    Figure Lengend Snippet: Effects of PMPP on β-catenin, Microphthalmia-associated transcription factor (MITF), tyrosinase (TYR) protein expression and TYR activity, and melanin content in B16 cells treated with GSK3β or Akt inhibitor. ( A ) The GSK3β inhibitor increased PMPP-induced β-catenin expression. β-catenin was detected in B16 cells pre-treated with or without BIO (5 μM) and incubated with PMPP (50 μM) or not. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 versus untreated cells, # p < 0.05 versus PMPP alone; ( B ) Effects of inhibitors on PMPP-induced MITF and TYR expression. The expressions of MITF and TYR were determined by Western blot in B16 cells treated with PMPP or not in the presence or absence of BIO (5 μM) or Akt inhibitor IV (1 μM) for 48 h. Data from the densitometric scanning of band intensities obtained from three separate experiments are presented as means ± SD. * p < 0.05, ** p < 0.01 versus MITF control, # p < 0.05, ## p < 0.01 versus TYR control. GSK3β inhibitor enhanced PMPP-induced TYR activity ( C ) and melanin content ( D ). The Akt inhibitor suppressed PMPP-induced TYR activity ( E ) and melanin content ( F ). B16 cells pre-incubated with inhibitors (BIO 5 μM, AKT inhibitor IV 1 μM) were further incubated with 50 μM PMPP, then followed by an additional incubation for 24 h (tyrosinase activity) or 48 h (melanin content). Each percentage value in the treated cells was calculated with respect to that in the untreated cells. Values are expressed as the mean ± SD of three separate experiments. ** p < 0.01, *** p < 0.001 compared with control; # p < 0.05, ### p < 0.001 compared with PMPP stimulation.

    Article Snippet: AKT (#5373), p-AKT s473 (#9271), GSK3β (#9832), p-GSK3β s9 (#9323), β-catenin (#8480), and β-actin (#3700) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). p-β-catenin s33 (sc-16743-R), antibodies against TYR (sc-7833), TRP-1 (sc-25543), and TRP-2 (sc-25544) were bought from Santa Cruz Technology (Dallas, TX, USA).

    Techniques: Expressing, Activity Assay, Incubation, Western Blot

    The possible mechanism of PMPP activating melanin content in B16 cells. PMPP increases the phosphorylation of Akt, and thus probably increases the non-activated GSK3β, which inhibits the phosphorylation of β-catenin and ceases its degradation via proteasomes. β-catenin accumulating in cytoplasm up-regulates MITF content through promoting transcription with LEF/TCF (Lymphoidenhancer factor/T cell factor) after coming into nucleus. The augmentation of β-catenin probably leads to the increases in the transcription of MITF and the TYR family and the subsequent induction of melanin synthesis.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: An Isoxazole Chalcone Derivative Enhances Melanogenesis in B16 Melanoma Cells via the Akt/GSK3β/β-Catenin Signaling Pathways

    doi: 10.3390/molecules22122077

    Figure Lengend Snippet: The possible mechanism of PMPP activating melanin content in B16 cells. PMPP increases the phosphorylation of Akt, and thus probably increases the non-activated GSK3β, which inhibits the phosphorylation of β-catenin and ceases its degradation via proteasomes. β-catenin accumulating in cytoplasm up-regulates MITF content through promoting transcription with LEF/TCF (Lymphoidenhancer factor/T cell factor) after coming into nucleus. The augmentation of β-catenin probably leads to the increases in the transcription of MITF and the TYR family and the subsequent induction of melanin synthesis.

    Article Snippet: AKT (#5373), p-AKT s473 (#9271), GSK3β (#9832), p-GSK3β s9 (#9323), β-catenin (#8480), and β-actin (#3700) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). p-β-catenin s33 (sc-16743-R), antibodies against TYR (sc-7833), TRP-1 (sc-25543), and TRP-2 (sc-25544) were bought from Santa Cruz Technology (Dallas, TX, USA).

    Techniques: