phospho gsk3β  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc phospho gsk3β
    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), <t>phospho-GSK3β</t> and GSK3β (panel C) antibodies as indicated.
    Phospho Gsk3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho gsk3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho gsk3β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "TGFβ-Stimulated MicroRNA-21 Utilizes PTEN to Orchestrate AKT/mTORC1 Signaling for Mesangial Cell Hypertrophy and Matrix Expansion"

    Article Title: TGFβ-Stimulated MicroRNA-21 Utilizes PTEN to Orchestrate AKT/mTORC1 Signaling for Mesangial Cell Hypertrophy and Matrix Expansion

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042316

    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), phospho-GSK3β and GSK3β (panel C) antibodies as indicated.
    Figure Legend Snippet: Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), phospho-GSK3β and GSK3β (panel C) antibodies as indicated.

    Techniques Used: Transfection, Plasmid Preparation, Incubation

    phospho gsk3β  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc phospho gsk3β
    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), <t>phospho-GSK3β</t> and GSK3β (panel C) antibodies as indicated.
    Phospho Gsk3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho gsk3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho gsk3β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "TGFβ-Stimulated MicroRNA-21 Utilizes PTEN to Orchestrate AKT/mTORC1 Signaling for Mesangial Cell Hypertrophy and Matrix Expansion"

    Article Title: TGFβ-Stimulated MicroRNA-21 Utilizes PTEN to Orchestrate AKT/mTORC1 Signaling for Mesangial Cell Hypertrophy and Matrix Expansion

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042316

    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), phospho-GSK3β and GSK3β (panel C) antibodies as indicated.
    Figure Legend Snippet: Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), phospho-GSK3β and GSK3β (panel C) antibodies as indicated.

    Techniques Used: Transfection, Plasmid Preparation, Incubation

    rabbit anti phospho gsk3 ser9  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc rabbit anti phospho gsk3 ser9
    Rabbit Anti Phospho Gsk3 Ser9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti phospho gsk3 ser9/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho gsk3 ser9 - by Bioz Stars, 2023-01
    97/100 stars

    Images

    rabbit anti glycogen synthase kinase gsk 3 β  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc rabbit anti glycogen synthase kinase gsk 3 β
    Rabbit Anti Glycogen Synthase Kinase Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti glycogen synthase kinase gsk 3 β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti glycogen synthase kinase gsk 3 β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    anti phosphorylated p gsk 3β antibodies  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc anti phosphorylated p gsk 3β antibodies
    Effect of cinnamaldehyde (Cin) on the level of total and phosphorylated (p)-glycogen synthase kinase <t>(GSK)-3β</t> in SHSY-5Y cells in the presence of beta amyloid (Aβ). The protein level was determined using Western blot analysis and normalized against untreated control cells. Data are presented as the mean ± standard error of the mean of four experiments. †: p<0.05 and †††: p<0.001 compared to the Aβ-treated cells; ₸: p<0.05 and ₸₸₸: p<0.001 compared to the vehicle-treated cells (control)
    Anti Phosphorylated P Gsk 3β Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phosphorylated p gsk 3β antibodies/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phosphorylated p gsk 3β antibodies - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "The neuroprotective mechanism of cinnamaldehyde against amyloid-β in neuronal SHSY5Y cell line: The role of N-methyl-D-aspartate, ryanodine, and adenosine receptors and glycogen synthase kinase-3β"

    Article Title: The neuroprotective mechanism of cinnamaldehyde against amyloid-β in neuronal SHSY5Y cell line: The role of N-methyl-D-aspartate, ryanodine, and adenosine receptors and glycogen synthase kinase-3β

    Journal: Avicenna Journal of Phytomedicine

    doi:

    Effect of cinnamaldehyde (Cin) on the level of total and phosphorylated (p)-glycogen synthase kinase (GSK)-3β in SHSY-5Y cells in the presence of beta amyloid (Aβ). The protein level was determined using Western blot analysis and normalized against untreated control cells. Data are presented as the mean ± standard error of the mean of four experiments. †: p<0.05 and †††: p<0.001 compared to the Aβ-treated cells; ₸: p<0.05 and ₸₸₸: p<0.001 compared to the vehicle-treated cells (control)
    Figure Legend Snippet: Effect of cinnamaldehyde (Cin) on the level of total and phosphorylated (p)-glycogen synthase kinase (GSK)-3β in SHSY-5Y cells in the presence of beta amyloid (Aβ). The protein level was determined using Western blot analysis and normalized against untreated control cells. Data are presented as the mean ± standard error of the mean of four experiments. †: p<0.05 and †††: p<0.001 compared to the Aβ-treated cells; ₸: p<0.05 and ₸₸₸: p<0.001 compared to the vehicle-treated cells (control)

    Techniques Used: Western Blot

    anti phospho gsk 3β ser9 rabbit pab  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc anti phospho gsk 3β ser9 rabbit pab
    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and <t>GSK-3β</t> were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.
    Anti Phospho Gsk 3β Ser9 Rabbit Pab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho gsk 3β ser9 rabbit pab/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho gsk 3β ser9 rabbit pab - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway"

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0053145

    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.
    Figure Legend Snippet: ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Luciferase, Construct, Infection, Western Blot, Transduction

    ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).
    Figure Legend Snippet: ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).

    Techniques Used: Inhibition, Activity Assay, Transfection, Dominant Negative Mutation, Construct, Plasmid Preparation, Transduction, Western Blot

    ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).
    Figure Legend Snippet: ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).

    Techniques Used: Over Expression, Transfection, Plasmid Preparation, Transduction, Western Blot

    ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).
    Figure Legend Snippet: ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).

    Techniques Used: Western Blot, Transfection, Plasmid Preparation, Transduction, Injection, Inhibition, Activity Assay

    anti gsk 3β rabbit pab  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc anti gsk 3β rabbit pab
    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and <t>GSK-3β</t> were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.
    Anti Gsk 3β Rabbit Pab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gsk 3β rabbit pab/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti gsk 3β rabbit pab - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway"

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0053145

    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.
    Figure Legend Snippet: ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Luciferase, Construct, Infection, Western Blot, Transduction

    ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).
    Figure Legend Snippet: ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).

    Techniques Used: Inhibition, Activity Assay, Transfection, Dominant Negative Mutation, Construct, Plasmid Preparation, Transduction, Western Blot

    ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).
    Figure Legend Snippet: ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).

    Techniques Used: Over Expression, Transfection, Plasmid Preparation, Transduction, Western Blot

    ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).
    Figure Legend Snippet: ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).

    Techniques Used: Western Blot, Transfection, Plasmid Preparation, Transduction, Injection, Inhibition, Activity Assay

    phospho gsk3β  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc phospho gsk3β
    Cells were transfected with Jnks (si1, si2, si3) or GFP (siGFP) siRNAs for 2 days and then exposed to cytokines (4 hrs). Western blot analysis was performed to determine ( A ) <t>phospho-GSK3β</t> or ( B ) phospho-FoxO3A and FoxO3A. Equal protein loading was assessed by blotting membranes with an antibody against tubulin. ( C ) Graphical presentations summarizing the effects of the different Jnk siRNAs vs siGFP in cytokine-treated cells; control values are set to 1. The data are the means±SD of three independent experiments. (**P<0.01) for cyto-Mix-siGFP vs cyto-Mix-si1, and cyto-Mix-si2. (***p<0.001) for cyto-Mix-siGFP vs cyto-Mix-si3. Significant differences were obtained for phospho-GSK3β in si1, si2, and si3 vs siGFP (***p<0.001).
    Phospho Gsk3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho gsk3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho gsk3β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "JNK3 Maintains Expression of the Insulin Receptor Substrate 2 (IRS2) in Insulin-Secreting Cells: Functional Consequences for Insulin Signaling"

    Article Title: JNK3 Maintains Expression of the Insulin Receptor Substrate 2 (IRS2) in Insulin-Secreting Cells: Functional Consequences for Insulin Signaling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0035997

    Cells were transfected with Jnks (si1, si2, si3) or GFP (siGFP) siRNAs for 2 days and then exposed to cytokines (4 hrs). Western blot analysis was performed to determine ( A ) phospho-GSK3β or ( B ) phospho-FoxO3A and FoxO3A. Equal protein loading was assessed by blotting membranes with an antibody against tubulin. ( C ) Graphical presentations summarizing the effects of the different Jnk siRNAs vs siGFP in cytokine-treated cells; control values are set to 1. The data are the means±SD of three independent experiments. (**P<0.01) for cyto-Mix-siGFP vs cyto-Mix-si1, and cyto-Mix-si2. (***p<0.001) for cyto-Mix-siGFP vs cyto-Mix-si3. Significant differences were obtained for phospho-GSK3β in si1, si2, and si3 vs siGFP (***p<0.001).
    Figure Legend Snippet: Cells were transfected with Jnks (si1, si2, si3) or GFP (siGFP) siRNAs for 2 days and then exposed to cytokines (4 hrs). Western blot analysis was performed to determine ( A ) phospho-GSK3β or ( B ) phospho-FoxO3A and FoxO3A. Equal protein loading was assessed by blotting membranes with an antibody against tubulin. ( C ) Graphical presentations summarizing the effects of the different Jnk siRNAs vs siGFP in cytokine-treated cells; control values are set to 1. The data are the means±SD of three independent experiments. (**P<0.01) for cyto-Mix-siGFP vs cyto-Mix-si1, and cyto-Mix-si2. (***p<0.001) for cyto-Mix-siGFP vs cyto-Mix-si3. Significant differences were obtained for phospho-GSK3β in si1, si2, and si3 vs siGFP (***p<0.001).

    Techniques Used: Transfection, Western Blot

    phospho gsk 3β  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc phospho gsk 3β
    A. Following treatment with TGF-β1 for 1 h, primary AEC exhibit marked phosphorylation of Akt at Ser437 by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated Akt phosphorylation. Membranes used for Western analysis were stripped and re-probed for total Akt to confirm equal protein loading and for normalization of p-Akt levels. *P <0.05 compared to TGF-β1; n = 3). B. Concomitant treatment with the PI3-K/Akt inhibitor LY294002 (0.5–3 µM) and TGF-β1 (2.5 ng/ml) attenuated Akt phosphorylation and subsequent induction of α-SMA by TGF-β1 in primary AEC. C. Quantitative analysis of α-SMA protein in primary AEC concomitantly treated with LY294002 (0.5–3 µM) and TGF-β1. *P <0.05 compared to TGF-β1; n≥3. D. Following treatment with TGF-β1 for 2 h, primary AEC exhibit marked phosphorylation of <t>GSK-3β</t> by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated GSK-3β phosphorylation. Membranes were re-probed for total GSK-3β to confirm equal protein loading and for normalization of pGSK-3β levels. *P <0.05 compared to TGF-β1; n = 3.
    Phospho Gsk 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho gsk 3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho gsk 3β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "Troglitazone Attenuates TGF-β1-Induced EMT in Alveolar Epithelial Cells via a PPARγ-Independent Mechanism"

    Article Title: Troglitazone Attenuates TGF-β1-Induced EMT in Alveolar Epithelial Cells via a PPARγ-Independent Mechanism

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0038827

    A. Following treatment with TGF-β1 for 1 h, primary AEC exhibit marked phosphorylation of Akt at Ser437 by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated Akt phosphorylation. Membranes used for Western analysis were stripped and re-probed for total Akt to confirm equal protein loading and for normalization of p-Akt levels. *P <0.05 compared to TGF-β1; n = 3). B. Concomitant treatment with the PI3-K/Akt inhibitor LY294002 (0.5–3 µM) and TGF-β1 (2.5 ng/ml) attenuated Akt phosphorylation and subsequent induction of α-SMA by TGF-β1 in primary AEC. C. Quantitative analysis of α-SMA protein in primary AEC concomitantly treated with LY294002 (0.5–3 µM) and TGF-β1. *P <0.05 compared to TGF-β1; n≥3. D. Following treatment with TGF-β1 for 2 h, primary AEC exhibit marked phosphorylation of GSK-3β by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated GSK-3β phosphorylation. Membranes were re-probed for total GSK-3β to confirm equal protein loading and for normalization of pGSK-3β levels. *P <0.05 compared to TGF-β1; n = 3.
    Figure Legend Snippet: A. Following treatment with TGF-β1 for 1 h, primary AEC exhibit marked phosphorylation of Akt at Ser437 by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated Akt phosphorylation. Membranes used for Western analysis were stripped and re-probed for total Akt to confirm equal protein loading and for normalization of p-Akt levels. *P <0.05 compared to TGF-β1; n = 3). B. Concomitant treatment with the PI3-K/Akt inhibitor LY294002 (0.5–3 µM) and TGF-β1 (2.5 ng/ml) attenuated Akt phosphorylation and subsequent induction of α-SMA by TGF-β1 in primary AEC. C. Quantitative analysis of α-SMA protein in primary AEC concomitantly treated with LY294002 (0.5–3 µM) and TGF-β1. *P <0.05 compared to TGF-β1; n≥3. D. Following treatment with TGF-β1 for 2 h, primary AEC exhibit marked phosphorylation of GSK-3β by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated GSK-3β phosphorylation. Membranes were re-probed for total GSK-3β to confirm equal protein loading and for normalization of pGSK-3β levels. *P <0.05 compared to TGF-β1; n = 3.

    Techniques Used: Western Blot

    phospho glycogen synthase kinase gsk 3β  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc phospho glycogen synthase kinase gsk 3β
    The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and <t>GSK-3β</t> in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.
    Phospho Glycogen Synthase Kinase Gsk 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho glycogen synthase kinase gsk 3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho glycogen synthase kinase gsk 3β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "Inhibition of Rho-associated coiled-coil containing protein kinase enhances the activation of epidermal growth factor receptor in pancreatic cancer cells"

    Article Title: Inhibition of Rho-associated coiled-coil containing protein kinase enhances the activation of epidermal growth factor receptor in pancreatic cancer cells

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-10-79

    The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and GSK-3β in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.
    Figure Legend Snippet: The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and GSK-3β in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.

    Techniques Used: Transfection, Negative Control, Western Blot, Software

    A schematic representation of the involvement of ROCK in the EGFR signaling in pancreatic cancer cells . First, EGF binds to the EGFR on the cell surface, and the EGFR undergoes dimerization and autophosphorylation at tyrosine residues. This triggers EGFR-related downstream signaling, such as activation of the MEK1/2-p44/p42 MAPK or Akt pathways, within 5 min of EGF stimulation (①). Around 5 min after the start of stimulation, EGF starts to induce the activation of RhoA and subsequently ROCK, as demonstrated by upregulation of the phosphorylation of cofilin and MLC (②). Inhibition experiments using Y27632 or siRNA indicated that activated ROCK cancels the persistent activation of the EGFR and the downstream pathways of MEK1/2 and Akt beginning at 5 min after the initiation of stimulation. Thus, ROCK functions to switch off the EGFR signaling that induces pancreatic cancer cell proliferation. EGF: epidermal growth factor, EGFR: EGF receptor, ROCK: Rho-associated coiled-coil containing protein kinase, MAPK: mitogen-activated protein kinase, GSK-3β: glycogen synthase kinase-3β, MLC, myosin light chain, P: phosphorylation, Y: tyrosine.
    Figure Legend Snippet: A schematic representation of the involvement of ROCK in the EGFR signaling in pancreatic cancer cells . First, EGF binds to the EGFR on the cell surface, and the EGFR undergoes dimerization and autophosphorylation at tyrosine residues. This triggers EGFR-related downstream signaling, such as activation of the MEK1/2-p44/p42 MAPK or Akt pathways, within 5 min of EGF stimulation (①). Around 5 min after the start of stimulation, EGF starts to induce the activation of RhoA and subsequently ROCK, as demonstrated by upregulation of the phosphorylation of cofilin and MLC (②). Inhibition experiments using Y27632 or siRNA indicated that activated ROCK cancels the persistent activation of the EGFR and the downstream pathways of MEK1/2 and Akt beginning at 5 min after the initiation of stimulation. Thus, ROCK functions to switch off the EGFR signaling that induces pancreatic cancer cell proliferation. EGF: epidermal growth factor, EGFR: EGF receptor, ROCK: Rho-associated coiled-coil containing protein kinase, MAPK: mitogen-activated protein kinase, GSK-3β: glycogen synthase kinase-3β, MLC, myosin light chain, P: phosphorylation, Y: tyrosine.

    Techniques Used: Activation Assay, Inhibition

    anti phospho gsk 3β  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Cell Signaling Technology Inc anti phospho gsk 3β
    Anti Phospho Gsk 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho gsk 3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho gsk 3β - by Bioz Stars, 2023-01
    97/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Cell Signaling Technology Inc phospho gsk3β
    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), <t>phospho-GSK3β</t> and GSK3β (panel C) antibodies as indicated.
    Phospho Gsk3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho gsk3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho gsk3β - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc rabbit anti phospho gsk3 ser9
    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), <t>phospho-GSK3β</t> and GSK3β (panel C) antibodies as indicated.
    Rabbit Anti Phospho Gsk3 Ser9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti phospho gsk3 ser9/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho gsk3 ser9 - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc rabbit anti glycogen synthase kinase gsk 3 β
    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), <t>phospho-GSK3β</t> and GSK3β (panel C) antibodies as indicated.
    Rabbit Anti Glycogen Synthase Kinase Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti glycogen synthase kinase gsk 3 β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti glycogen synthase kinase gsk 3 β - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc anti phosphorylated p gsk 3β antibodies
    Effect of cinnamaldehyde (Cin) on the level of total and phosphorylated (p)-glycogen synthase kinase <t>(GSK)-3β</t> in SHSY-5Y cells in the presence of beta amyloid (Aβ). The protein level was determined using Western blot analysis and normalized against untreated control cells. Data are presented as the mean ± standard error of the mean of four experiments. †: p<0.05 and †††: p<0.001 compared to the Aβ-treated cells; ₸: p<0.05 and ₸₸₸: p<0.001 compared to the vehicle-treated cells (control)
    Anti Phosphorylated P Gsk 3β Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phosphorylated p gsk 3β antibodies/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phosphorylated p gsk 3β antibodies - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc anti phospho gsk 3β ser9 rabbit pab
    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and <t>GSK-3β</t> were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.
    Anti Phospho Gsk 3β Ser9 Rabbit Pab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho gsk 3β ser9 rabbit pab/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho gsk 3β ser9 rabbit pab - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc anti gsk 3β rabbit pab
    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and <t>GSK-3β</t> were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.
    Anti Gsk 3β Rabbit Pab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gsk 3β rabbit pab/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti gsk 3β rabbit pab - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc phospho gsk 3β
    A. Following treatment with TGF-β1 for 1 h, primary AEC exhibit marked phosphorylation of Akt at Ser437 by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated Akt phosphorylation. Membranes used for Western analysis were stripped and re-probed for total Akt to confirm equal protein loading and for normalization of p-Akt levels. *P <0.05 compared to TGF-β1; n = 3). B. Concomitant treatment with the PI3-K/Akt inhibitor LY294002 (0.5–3 µM) and TGF-β1 (2.5 ng/ml) attenuated Akt phosphorylation and subsequent induction of α-SMA by TGF-β1 in primary AEC. C. Quantitative analysis of α-SMA protein in primary AEC concomitantly treated with LY294002 (0.5–3 µM) and TGF-β1. *P <0.05 compared to TGF-β1; n≥3. D. Following treatment with TGF-β1 for 2 h, primary AEC exhibit marked phosphorylation of <t>GSK-3β</t> by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated GSK-3β phosphorylation. Membranes were re-probed for total GSK-3β to confirm equal protein loading and for normalization of pGSK-3β levels. *P <0.05 compared to TGF-β1; n = 3.
    Phospho Gsk 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho gsk 3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho gsk 3β - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc phospho glycogen synthase kinase gsk 3β
    The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and <t>GSK-3β</t> in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.
    Phospho Glycogen Synthase Kinase Gsk 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho glycogen synthase kinase gsk 3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho glycogen synthase kinase gsk 3β - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Cell Signaling Technology Inc anti phospho gsk 3β
    The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and <t>GSK-3β</t> in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.
    Anti Phospho Gsk 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho gsk 3β/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho gsk 3β - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    Image Search Results


    Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), phospho-GSK3β and GSK3β (panel C) antibodies as indicated.

    Journal: PLoS ONE

    Article Title: TGFβ-Stimulated MicroRNA-21 Utilizes PTEN to Orchestrate AKT/mTORC1 Signaling for Mesangial Cell Hypertrophy and Matrix Expansion

    doi: 10.1371/journal.pone.0042316

    Figure Lengend Snippet: Mesangial cells were transfected with miR-21 Sponge or vector followed by incubation with 2 ng/ml TGFβ for 24 hours. Cell lysates were immunoblotted with PTEN, actin (panel A), phospho-Akt (Ser-473), phospho-Akt (Thr-308), Akt (panel B), phospho-GSK3β and GSK3β (panel C) antibodies as indicated.

    Article Snippet: Phospho-Akt (Ser473), phospho-Akt (thr-308), Akt, phospho-S6 kinase, S6 kinase, phospho-4EBP-1 (Thr-37/46) phospho-4EBP-1 (Ser-65) 4EBP-1, phospho-GSK3β, GSK3β, phospho-tuberin (Thr 1462), tuberin, phospho-PRAS40 (Thr 246), PRAS40, phospho-mTOR (Ser-2448) and mTOR antibodies were purchased from Cell Signaling, Boston, MA. siRNA pool of three oligonucleotides against PTEN mRNA, collagen II (α2) and PTEN antibodies were obtained from Santacruz, Delaware, CA.

    Techniques: Transfection, Plasmid Preparation, Incubation

    Effect of cinnamaldehyde (Cin) on the level of total and phosphorylated (p)-glycogen synthase kinase (GSK)-3β in SHSY-5Y cells in the presence of beta amyloid (Aβ). The protein level was determined using Western blot analysis and normalized against untreated control cells. Data are presented as the mean ± standard error of the mean of four experiments. †: p<0.05 and †††: p<0.001 compared to the Aβ-treated cells; ₸: p<0.05 and ₸₸₸: p<0.001 compared to the vehicle-treated cells (control)

    Journal: Avicenna Journal of Phytomedicine

    Article Title: The neuroprotective mechanism of cinnamaldehyde against amyloid-β in neuronal SHSY5Y cell line: The role of N-methyl-D-aspartate, ryanodine, and adenosine receptors and glycogen synthase kinase-3β

    doi:

    Figure Lengend Snippet: Effect of cinnamaldehyde (Cin) on the level of total and phosphorylated (p)-glycogen synthase kinase (GSK)-3β in SHSY-5Y cells in the presence of beta amyloid (Aβ). The protein level was determined using Western blot analysis and normalized against untreated control cells. Data are presented as the mean ± standard error of the mean of four experiments. †: p<0.05 and †††: p<0.001 compared to the Aβ-treated cells; ₸: p<0.05 and ₸₸₸: p<0.001 compared to the vehicle-treated cells (control)

    Article Snippet: The anti-actin, anti-GSK-3β, and anti-phosphorylated (p)-GSK-3β antibodies were obtained from Cell Signaling Technology ® (USA). the phosphate buffered solutions (PBS) was used as the solvent of cinnamaldehyde, dantrolene, adenosine, and NMDA.

    Techniques: Western Blot

    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Transfection, Expressing, Plasmid Preparation, Luciferase, Construct, Infection, Western Blot, Transduction

    ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Inhibition, Activity Assay, Transfection, Dominant Negative Mutation, Construct, Plasmid Preparation, Transduction, Western Blot

    ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Over Expression, Transfection, Plasmid Preparation, Transduction, Western Blot

    ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Western Blot, Transfection, Plasmid Preparation, Transduction, Injection, Inhibition, Activity Assay

    ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Effects of Tat on vIL-6 promoter activities in 293T cells. Cells were transfected with RTA expression plasmid ( With RTA ) or its control vector ( Without RTA ) and subsequently cotransfected with the firefly luciferase reporter construct pvIL-6-Luc (0.4 mg) and a Renilla luciferase construct pRL-TK (0.02 mg) following infection with Tat or Mock. Luciferase activities were measured, normalized to inner control, and presented as fold increase (n-fold). All data points were the averages of three independent experiments performed in triplicate. ( B ) Phosphorylation levels of PTEN, PI3K, AKT, and GSK-3β were analyzed by Western blot in 4E3 and T/V cells transduced by MOI 10 of Tat or Mock. Both 4E3 and T/V cells were starved with serum-free medium for 24 hours and followed by Tat or Mock transduction for 24 hours. The relative level of phosphorylation protein was determined by quantitative densitometry after normalized to corresponding non-phosphorylation protein. The relative level of phosphorylation protein in T/V+Mock group was considered to be 1 for comparison. ( C ) Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells on the CAM were analyzed by Western blot as above.

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Transfection, Expressing, Plasmid Preparation, Luciferase, Construct, Infection, Western Blot, Transduction

    ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Inhibition of PI3K activity suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 or T/V cells were transfected by the dominant-negative PI3K construct (PI3K-DN) and its control plasmid pSG5 for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 7). Two independent experiments were performed with similar results. ( B ) Inhibition of PI3K suppressed the enhanced effect of Tat on vIL-6-mediated tumorigenesis. 4E3 or T/V cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( C ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( D ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (C). Quantification of tumor weight was expressed as the mean ± SD (n = 5 to 6). ( E ) Inhibition of PI3K abrogated the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( F ) Inhibition of AKT abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (C). Collected cells were implanted onto the CAM. The phosphorylation level of GSK-3β in tumor tissues of the CAM model was analyzed by Western blot. ( G ) Inhibition of AKT activity restrained the promoted effect of Tat on vIL-6-mediated angiogenesis of endothelial cells. E6 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The representative angiogenic pictures and the quantification of blood vessels ( H ) on the CAM were shown. (n = 6 to 8). ( I ) Inhibition of AKT restrained the promoted effect of Tat on vIL-6-mediated tumorigenesis of endothelial cells. E6 cells were treated as in (G). The representative tumor pictures and the quantification of tumor weight ( J ) on the CAM were shown. The tumor weight was normalized to that of Matrigel alone and the data represents the mean ± SD (n = 6).

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Inhibition, Activity Assay, Transfection, Dominant Negative Mutation, Construct, Plasmid Preparation, Transduction, Western Blot

    ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by pPTEN and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). Two independent experiments were performed with similar results. ( B ) Overexpression of PTEN suppressed the enhanced effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were treated as in (A). Quantification of tumor weight was expressed as the mean ± SD (n = 5). ( C ) Overexpression of PTEN restrained the enhanced effect of Tat on phosphorylation of AKT and GSK-3β by vIL-6. 4E3 cells were treated as in (A). Collected cells were implanted onto the CAM. The phosphorylation levels of AKT and GSK-3β in tumor tissues of the CAM model were analyzed by Western blot. ( D ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated angiogenesis. 4E3 cells were transfected by the pS9A and its control plasmid pcDNA for 24 hours and followed by Tat or Mock transduction for another 24 hours. Collected cells were implanted onto the CAM. The quantification of blood vessels was expressed as the mean ± SD (n = 5 to 8). ( E ) Overexpression of GSK-3β decreased the promoted effect of Tat on vIL-6-mediated tumorigenesis. 4E3 cells were treated as in (D). Quantification of tumor weight was expressed as the mean ± SD (n = 5).

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Over Expression, Transfection, Plasmid Preparation, Transduction, Western Blot

    ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).

    Journal: PLoS ONE

    Article Title: HIV-1 Tat Promotes Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) vIL-6-Induced Angiogenesis and Tumorigenesis by Regulating PI3K/PTEN/AKT/GSK-3β Signaling Pathway

    doi: 10.1371/journal.pone.0053145

    Figure Lengend Snippet: ( A ) Tat augments vIL-6-induced phosphorylation of AKT and GSK-3β in nude mice. Phosphorylation AKT and GSK-3β in tumor tissues from 4E3 and T/V cells transduced by Tat and Mock in nude mice were analyzed by Western blot. ( B ) Plot of the volume of the tumors. 4E3 cells were transfected by the AKT-DN and its control plasmid pSRα for 24 hours and followed by Tat or Mock transduction for another 24 hours. Cells were injected s.c. into the left flank of the mice. The animals were monitored every day for the appearance of tumors until 30 days. Tumor size was estimated by two-dimensional caliper measurement. The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+AKT groups, respectively. △ P <.05 and △△ P <.01 for Student’s t -test versus Mock+pSRα groups, respectively. ns , not significant, t -test. ( C ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 30 after injections, and tumors were removed and weighed. Data reflect the mean ± SD. ( D ) Inhibition of AKT activity abolished the promoted effect of Tat on phosphorylation of GSK-3β by vIL-6. 4E3 cells were treated as in (B). The phosphorylation level of GSK-3β in tumor tissues of nude mice was analyzed by Western blot. ( E ) Plot of the volume of the tumors. 4E3 cells transduced with Tat or Mock were injected s.c. into mice for allograft formation. The mice received the treatments by intraperitoneal injection of LY294002 (the detailed procedure seen in ). The results are expressed as the mean ± SD (n = 5). * P <.05 and ** P <.01 for Student’s t -test versus Tat+LY294002 groups, respectively. △ P <.05 for Student’s t -test versus Mock+DMSO groups, respectively. ( F ) Column diagram of average tumor weight. Tumor-bearing mice were killed at day 40 after injections, and tumors were removed (Top) and weighed. Data reflect the mean ± SD (Bottom).

    Article Snippet: Anti-Flag rabbit pAb, anti-cyclin D1 rabbit mAb, anti-phospho-PTEN (Ser380) rabbit pAb, anti-PTEN rabbit pAb, anti-phospho-PI3K [p85(Tyr458)/p55(Tyr199)] rabbit pAb, anti-PI3K rabbit pAb, anti-phospho-AKT (Ser473) mouse mAb, anti-AKT mouse mAb, anti-phospho-GSK-3β (Ser9) rabbit pAb, anti-GSK-3β rabbit pAb, anti-CD31 and CD34 mouse mAbs were obtained from Cell Signaling Technologies (Danvers, MA).

    Techniques: Western Blot, Transfection, Plasmid Preparation, Transduction, Injection, Inhibition, Activity Assay

    A. Following treatment with TGF-β1 for 1 h, primary AEC exhibit marked phosphorylation of Akt at Ser437 by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated Akt phosphorylation. Membranes used for Western analysis were stripped and re-probed for total Akt to confirm equal protein loading and for normalization of p-Akt levels. *P <0.05 compared to TGF-β1; n = 3). B. Concomitant treatment with the PI3-K/Akt inhibitor LY294002 (0.5–3 µM) and TGF-β1 (2.5 ng/ml) attenuated Akt phosphorylation and subsequent induction of α-SMA by TGF-β1 in primary AEC. C. Quantitative analysis of α-SMA protein in primary AEC concomitantly treated with LY294002 (0.5–3 µM) and TGF-β1. *P <0.05 compared to TGF-β1; n≥3. D. Following treatment with TGF-β1 for 2 h, primary AEC exhibit marked phosphorylation of GSK-3β by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated GSK-3β phosphorylation. Membranes were re-probed for total GSK-3β to confirm equal protein loading and for normalization of pGSK-3β levels. *P <0.05 compared to TGF-β1; n = 3.

    Journal: PLoS ONE

    Article Title: Troglitazone Attenuates TGF-β1-Induced EMT in Alveolar Epithelial Cells via a PPARγ-Independent Mechanism

    doi: 10.1371/journal.pone.0038827

    Figure Lengend Snippet: A. Following treatment with TGF-β1 for 1 h, primary AEC exhibit marked phosphorylation of Akt at Ser437 by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated Akt phosphorylation. Membranes used for Western analysis were stripped and re-probed for total Akt to confirm equal protein loading and for normalization of p-Akt levels. *P <0.05 compared to TGF-β1; n = 3). B. Concomitant treatment with the PI3-K/Akt inhibitor LY294002 (0.5–3 µM) and TGF-β1 (2.5 ng/ml) attenuated Akt phosphorylation and subsequent induction of α-SMA by TGF-β1 in primary AEC. C. Quantitative analysis of α-SMA protein in primary AEC concomitantly treated with LY294002 (0.5–3 µM) and TGF-β1. *P <0.05 compared to TGF-β1; n≥3. D. Following treatment with TGF-β1 for 2 h, primary AEC exhibit marked phosphorylation of GSK-3β by Western analysis. Concomitant treatment with troglitazone (10 µM) and TGF-β1 (2.5 ng/mL) attenuated GSK-3β phosphorylation. Membranes were re-probed for total GSK-3β to confirm equal protein loading and for normalization of pGSK-3β levels. *P <0.05 compared to TGF-β1; n = 3.

    Article Snippet: Phospho-Akt (Ser473), total Akt, phospho-Smad2, total Smad2, phospho-Smad3, total Smad3, phospho-GSK-3β and total GSK-3β antibodies were purchased from Cell Signaling (Danvers, MA), and all secondary antibodies were obtained from Promega (Madison, WI).

    Techniques: Western Blot

    The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and GSK-3β in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.

    Journal: Molecular Cancer

    Article Title: Inhibition of Rho-associated coiled-coil containing protein kinase enhances the activation of epidermal growth factor receptor in pancreatic cancer cells

    doi: 10.1186/1476-4598-10-79

    Figure Lengend Snippet: The effects of Y27632 or ROCK1-siRNA on EGF-induced phosphorylation of MEK1/2, p44/p42 MAP kinase, Akt and GSK-3β in Panc1 cells . (A) The cells were pretreated with 3 μM Y27632 or vehicle for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for the indicated periods. (B) The cells were transfected with 10 nM of a negative control siRNA or siRNAs specifically targeting ROCK1 (#1) for 48 h, and then stimulated with 30 ng/ml of EGF or vehicle for 5 min. The cell lysates were then harvested and Western blotting were performed with antibodies against phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β and GAPDH. The intensities of protein bands in the Western blot analysis were determined by integrating the optical density over the band area using the NIH image software program. The intensity of each protein band was divided by the control (lane 1), and is shown above each panel.

    Article Snippet: Antibodies against phospho-cofilin, cofilin, phospho-myosin light chain (MLC), phospho-EGFR (Tyr1045 and Tyr1068), phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-glycogen synthase kinase (GSK)-3β, GSK-3β and ROCK1 were obtained from Cell Signaling, Inc. (Beverly, MA).

    Techniques: Transfection, Negative Control, Western Blot, Software

    A schematic representation of the involvement of ROCK in the EGFR signaling in pancreatic cancer cells . First, EGF binds to the EGFR on the cell surface, and the EGFR undergoes dimerization and autophosphorylation at tyrosine residues. This triggers EGFR-related downstream signaling, such as activation of the MEK1/2-p44/p42 MAPK or Akt pathways, within 5 min of EGF stimulation (①). Around 5 min after the start of stimulation, EGF starts to induce the activation of RhoA and subsequently ROCK, as demonstrated by upregulation of the phosphorylation of cofilin and MLC (②). Inhibition experiments using Y27632 or siRNA indicated that activated ROCK cancels the persistent activation of the EGFR and the downstream pathways of MEK1/2 and Akt beginning at 5 min after the initiation of stimulation. Thus, ROCK functions to switch off the EGFR signaling that induces pancreatic cancer cell proliferation. EGF: epidermal growth factor, EGFR: EGF receptor, ROCK: Rho-associated coiled-coil containing protein kinase, MAPK: mitogen-activated protein kinase, GSK-3β: glycogen synthase kinase-3β, MLC, myosin light chain, P: phosphorylation, Y: tyrosine.

    Journal: Molecular Cancer

    Article Title: Inhibition of Rho-associated coiled-coil containing protein kinase enhances the activation of epidermal growth factor receptor in pancreatic cancer cells

    doi: 10.1186/1476-4598-10-79

    Figure Lengend Snippet: A schematic representation of the involvement of ROCK in the EGFR signaling in pancreatic cancer cells . First, EGF binds to the EGFR on the cell surface, and the EGFR undergoes dimerization and autophosphorylation at tyrosine residues. This triggers EGFR-related downstream signaling, such as activation of the MEK1/2-p44/p42 MAPK or Akt pathways, within 5 min of EGF stimulation (①). Around 5 min after the start of stimulation, EGF starts to induce the activation of RhoA and subsequently ROCK, as demonstrated by upregulation of the phosphorylation of cofilin and MLC (②). Inhibition experiments using Y27632 or siRNA indicated that activated ROCK cancels the persistent activation of the EGFR and the downstream pathways of MEK1/2 and Akt beginning at 5 min after the initiation of stimulation. Thus, ROCK functions to switch off the EGFR signaling that induces pancreatic cancer cell proliferation. EGF: epidermal growth factor, EGFR: EGF receptor, ROCK: Rho-associated coiled-coil containing protein kinase, MAPK: mitogen-activated protein kinase, GSK-3β: glycogen synthase kinase-3β, MLC, myosin light chain, P: phosphorylation, Y: tyrosine.

    Article Snippet: Antibodies against phospho-cofilin, cofilin, phospho-myosin light chain (MLC), phospho-EGFR (Tyr1045 and Tyr1068), phospho-MEK1/2, MEK1/2, phospho-p44/p42 MAP kinase, p44/p42 MAP kinase, phospho-Akt, Akt, phospho-glycogen synthase kinase (GSK)-3β, GSK-3β and ROCK1 were obtained from Cell Signaling, Inc. (Beverly, MA).

    Techniques: Activation Assay, Inhibition