p gsk3 β ser9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk3 β ser9
    Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, <t>p-GSK3</t> β <t>(Ser9),</t> and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.
    P Gsk3 β Ser9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Leptin Promotes HTR-8/SVneo Cell Invasion via the Crosstalk between MTA1/WNT and PI3K/AKT Pathways"

    Article Title: Leptin Promotes HTR-8/SVneo Cell Invasion via the Crosstalk between MTA1/WNT and PI3K/AKT Pathways

    Journal: Disease Markers

    doi: 10.1155/2022/7052176

    Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, p-GSK3 β (Ser9), and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.
    Figure Legend Snippet: Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, p-GSK3 β (Ser9), and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.

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

    anti p gsk 3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p gsk 3 β
    Effects of HA-ADT on the apoptotic level and <t>AKT/GSK-3</t> β / β -catenin pathway in human HCC cells. (a) TUNEL staining was used to detect the apoptotic level (original magnification, 100x). (b) The apoptotic index was counted as the ratio of TUNEL positive cells to total cells. (c) Flow cytometry assay was adopted to detect apoptosis. (d) Cell apoptosis distribution was analyzed. (e) Western blotting was used to determine the protein levels of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, and p- β -catenin. β -actin was adopted as the internal control. (f) The density was analyzed. All data are shown as the mean ± SEM of three independent experiments; ∗ P < 0.05, ∗∗ P < 0.01 vs. control group; △ P < 0.05, △△ P < 0.01 vs. NaHS group; ## P < 0.01 vs. GYY4137 group.
    Anti P Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A Water-Soluble Hydrogen Sulfide Donor Suppresses the Growth of Hepatocellular Carcinoma via Inhibiting the AKT/GSK-3 β / β -Catenin and TGF- β /Smad2/3 Signaling Pathways"

    Article Title: A Water-Soluble Hydrogen Sulfide Donor Suppresses the Growth of Hepatocellular Carcinoma via Inhibiting the AKT/GSK-3 β / β -Catenin and TGF- β /Smad2/3 Signaling Pathways

    Journal: Journal of Oncology

    doi: 10.1155/2023/8456852

    Effects of HA-ADT on the apoptotic level and AKT/GSK-3 β / β -catenin pathway in human HCC cells. (a) TUNEL staining was used to detect the apoptotic level (original magnification, 100x). (b) The apoptotic index was counted as the ratio of TUNEL positive cells to total cells. (c) Flow cytometry assay was adopted to detect apoptosis. (d) Cell apoptosis distribution was analyzed. (e) Western blotting was used to determine the protein levels of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, and p- β -catenin. β -actin was adopted as the internal control. (f) The density was analyzed. All data are shown as the mean ± SEM of three independent experiments; ∗ P < 0.05, ∗∗ P < 0.01 vs. control group; △ P < 0.05, △△ P < 0.01 vs. NaHS group; ## P < 0.01 vs. GYY4137 group.
    Figure Legend Snippet: Effects of HA-ADT on the apoptotic level and AKT/GSK-3 β / β -catenin pathway in human HCC cells. (a) TUNEL staining was used to detect the apoptotic level (original magnification, 100x). (b) The apoptotic index was counted as the ratio of TUNEL positive cells to total cells. (c) Flow cytometry assay was adopted to detect apoptosis. (d) Cell apoptosis distribution was analyzed. (e) Western blotting was used to determine the protein levels of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, and p- β -catenin. β -actin was adopted as the internal control. (f) The density was analyzed. All data are shown as the mean ± SEM of three independent experiments; ∗ P < 0.05, ∗∗ P < 0.01 vs. control group; △ P < 0.05, △△ P < 0.01 vs. NaHS group; ## P < 0.01 vs. GYY4137 group.

    Techniques Used: TUNEL Assay, Staining, Flow Cytometry, Western Blot

    p gsk3 β ser9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk3 β ser9
    Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, <t>p-GSK3</t> β <t>(Ser9),</t> and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.
    P Gsk3 β Ser9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Leptin Promotes HTR-8/SVneo Cell Invasion via the Crosstalk between MTA1/WNT and PI3K/AKT Pathways"

    Article Title: Leptin Promotes HTR-8/SVneo Cell Invasion via the Crosstalk between MTA1/WNT and PI3K/AKT Pathways

    Journal: Disease Markers

    doi: 10.1155/2022/7052176

    Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, p-GSK3 β (Ser9), and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.
    Figure Legend Snippet: Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, p-GSK3 β (Ser9), and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.

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

    p gsk3β ser9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk3β ser9
    A , B The activation level of <t>AKT/GSK3β</t> pathway was detected by western blot in vitro and in vivo. C , D The expression levels of EMT marker molecules (N-cadherin, E-cadherin and TGFβ1), inflammatory pathway protein molecules (NLRP3, Cleaved Caspase1/Caspase1) and fibrosis marker molecules (COL-1, COL-3 and α-SMA) was detected by western blot. E Inflammatory factor IL-1β and inflammatory chemokine CXCL2 the supernatant of alveolar epithelial cells were detected by ELISA. Data were expressed as the mean ± SD, n = 3. ** P < 0.01 and *** P < 0.001. F The migration and invasion abilities of alveolar epithelial cells were detected by scratch healing assay and transwell assay.
    P Gsk3β Ser9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Coal dust nanoparticles induced pulmonary fibrosis by promoting inflammation and epithelial-mesenchymal transition via the NF-κB/NLRP3 pathway driven by IGF1/ROS-mediated AKT/GSK3β signals"

    Article Title: Coal dust nanoparticles induced pulmonary fibrosis by promoting inflammation and epithelial-mesenchymal transition via the NF-κB/NLRP3 pathway driven by IGF1/ROS-mediated AKT/GSK3β signals

    Journal: Cell Death Discovery

    doi: 10.1038/s41420-022-01291-z

    A , B The activation level of AKT/GSK3β pathway was detected by western blot in vitro and in vivo. C , D The expression levels of EMT marker molecules (N-cadherin, E-cadherin and TGFβ1), inflammatory pathway protein molecules (NLRP3, Cleaved Caspase1/Caspase1) and fibrosis marker molecules (COL-1, COL-3 and α-SMA) was detected by western blot. E Inflammatory factor IL-1β and inflammatory chemokine CXCL2 the supernatant of alveolar epithelial cells were detected by ELISA. Data were expressed as the mean ± SD, n = 3. ** P < 0.01 and *** P < 0.001. F The migration and invasion abilities of alveolar epithelial cells were detected by scratch healing assay and transwell assay.
    Figure Legend Snippet: A , B The activation level of AKT/GSK3β pathway was detected by western blot in vitro and in vivo. C , D The expression levels of EMT marker molecules (N-cadherin, E-cadherin and TGFβ1), inflammatory pathway protein molecules (NLRP3, Cleaved Caspase1/Caspase1) and fibrosis marker molecules (COL-1, COL-3 and α-SMA) was detected by western blot. E Inflammatory factor IL-1β and inflammatory chemokine CXCL2 the supernatant of alveolar epithelial cells were detected by ELISA. Data were expressed as the mean ± SD, n = 3. ** P < 0.01 and *** P < 0.001. F The migration and invasion abilities of alveolar epithelial cells were detected by scratch healing assay and transwell assay.

    Techniques Used: Activation Assay, Western Blot, In Vitro, In Vivo, Expressing, Marker, Enzyme-linked Immunosorbent Assay, Migration, Transwell Assay

    p gsk3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk3 β
    Protein expression in GA muscle samples from rats treated with n-3 PUFA or vehicle for 40 days and dexamethasone or vehicle (last 10 days). Relative optical density of total Akt (tAkt) and phosphorylated Akt (P-Akt) ((a) and (c)), total <t>GSK3</t> <t>β</t> (tGSK3 β ) and phosphorylated GSK3 β (P-GSK3 β ) ((b) and (d)), and tAkt/P-Akt and tGSK3 β /P-GSK3 β ratios ((e) and (f), resp.). Bands representing tAkt, P-Akt, tGSK3, and P-GSK3 β are illustrated in (g). α -Tubulin (55 kDa) was used as control. The numbers represent the mean ± S.E.M. CT = control group ( n = 6), DX = dexamethasone group ( n = 5), n-3 = n-3 PUFA group ( n = 6), and DX + n-3 = dexamethasone + n-3 PUFA group ( n = 6). Values represent means ± S.E.M., analyzed by Student's t -test, # P < 0.05.
    P Gsk3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "The Effects of Omega-3 Fatty Acid Supplementation on Dexamethasone-Induced Muscle Atrophy"

    Article Title: The Effects of Omega-3 Fatty Acid Supplementation on Dexamethasone-Induced Muscle Atrophy

    Journal: BioMed Research International

    doi: 10.1155/2014/961438

    Protein expression in GA muscle samples from rats treated with n-3 PUFA or vehicle for 40 days and dexamethasone or vehicle (last 10 days). Relative optical density of total Akt (tAkt) and phosphorylated Akt (P-Akt) ((a) and (c)), total GSK3 β (tGSK3 β ) and phosphorylated GSK3 β (P-GSK3 β ) ((b) and (d)), and tAkt/P-Akt and tGSK3 β /P-GSK3 β ratios ((e) and (f), resp.). Bands representing tAkt, P-Akt, tGSK3, and P-GSK3 β are illustrated in (g). α -Tubulin (55 kDa) was used as control. The numbers represent the mean ± S.E.M. CT = control group ( n = 6), DX = dexamethasone group ( n = 5), n-3 = n-3 PUFA group ( n = 6), and DX + n-3 = dexamethasone + n-3 PUFA group ( n = 6). Values represent means ± S.E.M., analyzed by Student's t -test, # P < 0.05.
    Figure Legend Snippet: Protein expression in GA muscle samples from rats treated with n-3 PUFA or vehicle for 40 days and dexamethasone or vehicle (last 10 days). Relative optical density of total Akt (tAkt) and phosphorylated Akt (P-Akt) ((a) and (c)), total GSK3 β (tGSK3 β ) and phosphorylated GSK3 β (P-GSK3 β ) ((b) and (d)), and tAkt/P-Akt and tGSK3 β /P-GSK3 β ratios ((e) and (f), resp.). Bands representing tAkt, P-Akt, tGSK3, and P-GSK3 β are illustrated in (g). α -Tubulin (55 kDa) was used as control. The numbers represent the mean ± S.E.M. CT = control group ( n = 6), DX = dexamethasone group ( n = 5), n-3 = n-3 PUFA group ( n = 6), and DX + n-3 = dexamethasone + n-3 PUFA group ( n = 6). Values represent means ± S.E.M., analyzed by Student's t -test, # P < 0.05.

    Techniques Used: Expressing

    p gsk 3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk 3 β
    Effects of DSG and 5-MOP on the expression of proteins in PI3K/Akt pathways involving ER α . Representative protein levels for (a) p-ER (Tyr-537)/ER α , (b) p-Src (Tyr-416)/Src, (c) PI3Kp85/ β -actin, (d) p-Akt (Ser473)/Akt, and (e) <t>p-GSK-3</t> β <t>(Ser9)/GSK-3</t> β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. ## P < 0.01: significance from DSG-treated group.
    P Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Diosgenin and 5-Methoxypsoralen Ameliorate Insulin Resistance through ER- α /PI3K/Akt-Signaling Pathways in HepG2 Cells"

    Article Title: Diosgenin and 5-Methoxypsoralen Ameliorate Insulin Resistance through ER- α /PI3K/Akt-Signaling Pathways in HepG2 Cells

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2016/7493694

    Effects of DSG and 5-MOP on the expression of proteins in PI3K/Akt pathways involving ER α . Representative protein levels for (a) p-ER (Tyr-537)/ER α , (b) p-Src (Tyr-416)/Src, (c) PI3Kp85/ β -actin, (d) p-Akt (Ser473)/Akt, and (e) p-GSK-3 β (Ser9)/GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. ## P < 0.01: significance from DSG-treated group.
    Figure Legend Snippet: Effects of DSG and 5-MOP on the expression of proteins in PI3K/Akt pathways involving ER α . Representative protein levels for (a) p-ER (Tyr-537)/ER α , (b) p-Src (Tyr-416)/Src, (c) PI3Kp85/ β -actin, (d) p-Akt (Ser473)/Akt, and (e) p-GSK-3 β (Ser9)/GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. ## P < 0.01: significance from DSG-treated group.

    Techniques Used: Expressing

    Effects of DSG and 5-MOP on gene expression at the transcriptional level in PI3K/Akt pathways involving ER α . Representative mRNA levels for (a) ER α , (b) Src, (c) PI3K p85, (d) Akt, and (e) GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. # P < 0.05: significance from DSG-treated group.
    Figure Legend Snippet: Effects of DSG and 5-MOP on gene expression at the transcriptional level in PI3K/Akt pathways involving ER α . Representative mRNA levels for (a) ER α , (b) Src, (c) PI3K p85, (d) Akt, and (e) GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. # P < 0.05: significance from DSG-treated group.

    Techniques Used: Expressing

    p gsk3β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk3β
    A, Western blots of MEK1/2, ERK1/2, p38, and JNK phosphorylation and their total protein levels 28 days after surgery in WT and KO mice. GAPDH was used as the sample loading control. (n = 6). Up, representative blots. Down, quantitative results. B, Western blots of <t>Akt,GSK3β,</t> and mTOR phosphorylation and their total protein levels 28 days after surgery. GAPDH was used as the sample loading control (n = 6). Up, representative blots. Down, quantitative results. For A and B, Data represent as the means ± SEM *P<0.01 vs. corresponding sham; #P<0.01 vs. WT/AB after AB.
    P Gsk3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Augmented Cardiac Hypertrophy in Response to Pressure Overload in Mice Lacking ELTD1"

    Article Title: Augmented Cardiac Hypertrophy in Response to Pressure Overload in Mice Lacking ELTD1

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0035779

    A, Western blots of MEK1/2, ERK1/2, p38, and JNK phosphorylation and their total protein levels 28 days after surgery in WT and KO mice. GAPDH was used as the sample loading control. (n = 6). Up, representative blots. Down, quantitative results. B, Western blots of Akt,GSK3β, and mTOR phosphorylation and their total protein levels 28 days after surgery. GAPDH was used as the sample loading control (n = 6). Up, representative blots. Down, quantitative results. For A and B, Data represent as the means ± SEM *P<0.01 vs. corresponding sham; #P<0.01 vs. WT/AB after AB.
    Figure Legend Snippet: A, Western blots of MEK1/2, ERK1/2, p38, and JNK phosphorylation and their total protein levels 28 days after surgery in WT and KO mice. GAPDH was used as the sample loading control. (n = 6). Up, representative blots. Down, quantitative results. B, Western blots of Akt,GSK3β, and mTOR phosphorylation and their total protein levels 28 days after surgery. GAPDH was used as the sample loading control (n = 6). Up, representative blots. Down, quantitative results. For A and B, Data represent as the means ± SEM *P<0.01 vs. corresponding sham; #P<0.01 vs. WT/AB after AB.

    Techniques Used: Western Blot

    p gsk 3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk 3 β
    Comparison of protein kinase B (AKT) and glycogen synthase kinase-3 β <t>(GSK-3</t> β ) phosphorylation levels among different groups. The five groups were treated as above 3. (a) Representative Western blot analysis of AKT, P-AKT Ser473 , GSK-3 β , and P-GSK-3 β Ser9 protein of BRL-3A cells stimulated by LPS. The total AKT and GSK-3 β were used as a control. (b) Quantitative analysis of P-AKT Ser473 /AKT and P-GSK-3 β Ser9 /GSK-3 β . Data were mean ± SEM ( n = 3). ∗ P < 0.05 versus control group. # P < 0.05 versus LPS group.
    P Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Role of TLR4-Mediated PI3K/AKT/GSK-3 β Signaling Pathway in Apoptosis of Rat Hepatocytes"

    Article Title: Role of TLR4-Mediated PI3K/AKT/GSK-3 β Signaling Pathway in Apoptosis of Rat Hepatocytes

    Journal: BioMed Research International

    doi: 10.1155/2015/631326

    Comparison of protein kinase B (AKT) and glycogen synthase kinase-3 β (GSK-3 β ) phosphorylation levels among different groups. The five groups were treated as above 3. (a) Representative Western blot analysis of AKT, P-AKT Ser473 , GSK-3 β , and P-GSK-3 β Ser9 protein of BRL-3A cells stimulated by LPS. The total AKT and GSK-3 β were used as a control. (b) Quantitative analysis of P-AKT Ser473 /AKT and P-GSK-3 β Ser9 /GSK-3 β . Data were mean ± SEM ( n = 3). ∗ P < 0.05 versus control group. # P < 0.05 versus LPS group.
    Figure Legend Snippet: Comparison of protein kinase B (AKT) and glycogen synthase kinase-3 β (GSK-3 β ) phosphorylation levels among different groups. The five groups were treated as above 3. (a) Representative Western blot analysis of AKT, P-AKT Ser473 , GSK-3 β , and P-GSK-3 β Ser9 protein of BRL-3A cells stimulated by LPS. The total AKT and GSK-3 β were used as a control. (b) Quantitative analysis of P-AKT Ser473 /AKT and P-GSK-3 β Ser9 /GSK-3 β . Data were mean ± SEM ( n = 3). ∗ P < 0.05 versus control group. # P < 0.05 versus LPS group.

    Techniques Used: Western Blot

    TLR4 inhibition, GSK-3 β inhibition, and AKT inhibition affect GSK-3 β translocation induced by LPS. The five groups were treated as above 4. GSK-3 β translocated to the nucleus in LPS group (white arrow) and LY294002 + LPS group (yellow arrows). BRL-3A cells were incubated with GSK-3 β antibody. Nucleus was visualized using Hoechst 33342 (blue). Immunofluorescence microscopy was used to detect the location of GSK-3 β (green). Scale bars = 10 μ m.
    Figure Legend Snippet: TLR4 inhibition, GSK-3 β inhibition, and AKT inhibition affect GSK-3 β translocation induced by LPS. The five groups were treated as above 4. GSK-3 β translocated to the nucleus in LPS group (white arrow) and LY294002 + LPS group (yellow arrows). BRL-3A cells were incubated with GSK-3 β antibody. Nucleus was visualized using Hoechst 33342 (blue). Immunofluorescence microscopy was used to detect the location of GSK-3 β (green). Scale bars = 10 μ m.

    Techniques Used: Inhibition, Translocation Assay, Incubation, Immunofluorescence, Microscopy

    TLR4/PI3K/AKT/GSK-3 β signaling pathway inhibition affects production of Bax, Bcl-2, and active caspase-3. The five groups were treated as above 4. (a) The mRNA expression of Bax/Bcl-2 and caspase-3 was detected by RT-qPCR. (b) Quantification (relative density) of the intensity of staining of active caspase-3, Bax, and Bcl-2 protein detected by Western blot. (c) Representative Western blot of active caspase-3, Bax, and Bcl-2 protein from BRL-3A cells. β -actin was used as an internal control. Data were mean ± SEM ( n = 3). ∗ P < 0.05 versus control group. # P < 0.05 versus LPS group.
    Figure Legend Snippet: TLR4/PI3K/AKT/GSK-3 β signaling pathway inhibition affects production of Bax, Bcl-2, and active caspase-3. The five groups were treated as above 4. (a) The mRNA expression of Bax/Bcl-2 and caspase-3 was detected by RT-qPCR. (b) Quantification (relative density) of the intensity of staining of active caspase-3, Bax, and Bcl-2 protein detected by Western blot. (c) Representative Western blot of active caspase-3, Bax, and Bcl-2 protein from BRL-3A cells. β -actin was used as an internal control. Data were mean ± SEM ( n = 3). ∗ P < 0.05 versus control group. # P < 0.05 versus LPS group.

    Techniques Used: Inhibition, Expressing, Quantitative RT-PCR, Staining, Western Blot

    p gsk 3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk 3 β
    P Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p gsk 3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk 3 β
    ZSD induced the apoptosis of H1299 cells partially by regulating <t>AKT/GSK-3</t> β / β -catenin signaling. (a) ZSD induced the apoptosis of H1299 cells after treatment with ZSD (4 mg/mL) for 12 h, 24 h, and 48 h. (b) ZSD induced apoptosis of H1299 cells in a dose-dependent manner. (c) Pathway enrichment analysis of the genes in Human Signal Transduction Pathway Finder PCR Array. (d) The mRNA expression of AKT/GSK-3 β / β -catenin signal cascades in H1299 cells after treatment with ZSD (4 mg/mL). ∗ p < 0.05 and ∗∗ p < 0.01 vs. the control group.
    P Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Zi Shen Decoction Inhibits Growth and Metastasis of Lung Cancer via Regulating the AKT/GSK-3 β / β -Catenin Pathway"

    Article Title: Zi Shen Decoction Inhibits Growth and Metastasis of Lung Cancer via Regulating the AKT/GSK-3 β / β -Catenin Pathway

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2021/6685282

    ZSD induced the apoptosis of H1299 cells partially by regulating AKT/GSK-3 β / β -catenin signaling. (a) ZSD induced the apoptosis of H1299 cells after treatment with ZSD (4 mg/mL) for 12 h, 24 h, and 48 h. (b) ZSD induced apoptosis of H1299 cells in a dose-dependent manner. (c) Pathway enrichment analysis of the genes in Human Signal Transduction Pathway Finder PCR Array. (d) The mRNA expression of AKT/GSK-3 β / β -catenin signal cascades in H1299 cells after treatment with ZSD (4 mg/mL). ∗ p < 0.05 and ∗∗ p < 0.01 vs. the control group.
    Figure Legend Snippet: ZSD induced the apoptosis of H1299 cells partially by regulating AKT/GSK-3 β / β -catenin signaling. (a) ZSD induced the apoptosis of H1299 cells after treatment with ZSD (4 mg/mL) for 12 h, 24 h, and 48 h. (b) ZSD induced apoptosis of H1299 cells in a dose-dependent manner. (c) Pathway enrichment analysis of the genes in Human Signal Transduction Pathway Finder PCR Array. (d) The mRNA expression of AKT/GSK-3 β / β -catenin signal cascades in H1299 cells after treatment with ZSD (4 mg/mL). ∗ p < 0.05 and ∗∗ p < 0.01 vs. the control group.

    Techniques Used: Transduction, Expressing

    ZSD inhibited the AKT/GSK-3 β / β -catenin pathway in lung cancer cells and tumor tissues. (a) Western blot analysis was used to assess the expression of multiple proteins involved in the AKT/GSK-3 β / β -catenin pathway in lung cancer cells after ZSD treatment for 12 h, 24 h, and 48 h. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the control group. (b) Western blots showing the protein expression of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, cleaved caspase-3, and Bax and Bcl-2 in isolated tumor tissues. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the model group.
    Figure Legend Snippet: ZSD inhibited the AKT/GSK-3 β / β -catenin pathway in lung cancer cells and tumor tissues. (a) Western blot analysis was used to assess the expression of multiple proteins involved in the AKT/GSK-3 β / β -catenin pathway in lung cancer cells after ZSD treatment for 12 h, 24 h, and 48 h. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the control group. (b) Western blots showing the protein expression of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, cleaved caspase-3, and Bax and Bcl-2 in isolated tumor tissues. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the model group.

    Techniques Used: Western Blot, Expressing, Isolation

    The AKT/GSK-3 β / β -catenin pathway medicated anticancer effect of ZSD in lung cancer cells. H1299 cells were treated with ZSD and/or SC-79 for 24 h. (a) Representative western blots and quantitative analysis of the AKT/GSK-3 β / β -catenin pathway related proteins. (b) The mobility of H1299 cells was detected by wound-healing assay. (c) The migration and invasion abilities of H1299 cells were detected by transwell assays. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the control group.
    Figure Legend Snippet: The AKT/GSK-3 β / β -catenin pathway medicated anticancer effect of ZSD in lung cancer cells. H1299 cells were treated with ZSD and/or SC-79 for 24 h. (a) Representative western blots and quantitative analysis of the AKT/GSK-3 β / β -catenin pathway related proteins. (b) The mobility of H1299 cells was detected by wound-healing assay. (c) The migration and invasion abilities of H1299 cells were detected by transwell assays. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the control group.

    Techniques Used: Western Blot, Wound Healing Assay, Migration

    p gsk 3β ser9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p gsk 3β ser9
    A and B, The phosphorylated and total protein levels of Akt, <t>GSK-3β,</t> mTOR, eIF4E, and 4E-BP1 in WT and Nek6 KO mice 4 weeks after TAC or sham surgery. Nek6 deficiency augments the activation of Akt signaling. A, representative western blots (duplicate lanes represent two different heart samples); B, quantitative results. * P <0.05 vs . WT/sham; # P <0.05 vs . WT/TAC. C and D, phosphorylated and total protein expression levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in H9c2 cells treated with Ang II for the indicated times with or without (control) the overexpression of Nek6. Nek6 overexpression attenuated the activation of Akt signaling. C, representative western blots; D, quantitative results. * P <0.05 vs. the control group at the 0 time point; # P <0.05 vs . the control group at the same time point.
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    1) Product Images from "Never in Mitosis Gene A Related Kinase-6 Attenuates Pressure Overload-Induced Activation of the Protein Kinase B Pathway and Cardiac Hypertrophy"

    Article Title: Never in Mitosis Gene A Related Kinase-6 Attenuates Pressure Overload-Induced Activation of the Protein Kinase B Pathway and Cardiac Hypertrophy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0096095

    A and B, The phosphorylated and total protein levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in WT and Nek6 KO mice 4 weeks after TAC or sham surgery. Nek6 deficiency augments the activation of Akt signaling. A, representative western blots (duplicate lanes represent two different heart samples); B, quantitative results. * P <0.05 vs . WT/sham; # P <0.05 vs . WT/TAC. C and D, phosphorylated and total protein expression levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in H9c2 cells treated with Ang II for the indicated times with or without (control) the overexpression of Nek6. Nek6 overexpression attenuated the activation of Akt signaling. C, representative western blots; D, quantitative results. * P <0.05 vs. the control group at the 0 time point; # P <0.05 vs . the control group at the same time point.
    Figure Legend Snippet: A and B, The phosphorylated and total protein levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in WT and Nek6 KO mice 4 weeks after TAC or sham surgery. Nek6 deficiency augments the activation of Akt signaling. A, representative western blots (duplicate lanes represent two different heart samples); B, quantitative results. * P <0.05 vs . WT/sham; # P <0.05 vs . WT/TAC. C and D, phosphorylated and total protein expression levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in H9c2 cells treated with Ang II for the indicated times with or without (control) the overexpression of Nek6. Nek6 overexpression attenuated the activation of Akt signaling. C, representative western blots; D, quantitative results. * P <0.05 vs. the control group at the 0 time point; # P <0.05 vs . the control group at the same time point.

    Techniques Used: Activation Assay, Western Blot, Expressing, Over Expression

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    Cell Signaling Technology Inc p gsk3 β ser9
    Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, <t>p-GSK3</t> β <t>(Ser9),</t> and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.
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    Cell Signaling Technology Inc anti p gsk 3 β
    Effects of HA-ADT on the apoptotic level and <t>AKT/GSK-3</t> β / β -catenin pathway in human HCC cells. (a) TUNEL staining was used to detect the apoptotic level (original magnification, 100x). (b) The apoptotic index was counted as the ratio of TUNEL positive cells to total cells. (c) Flow cytometry assay was adopted to detect apoptosis. (d) Cell apoptosis distribution was analyzed. (e) Western blotting was used to determine the protein levels of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, and p- β -catenin. β -actin was adopted as the internal control. (f) The density was analyzed. All data are shown as the mean ± SEM of three independent experiments; ∗ P < 0.05, ∗∗ P < 0.01 vs. control group; △ P < 0.05, △△ P < 0.01 vs. NaHS group; ## P < 0.01 vs. GYY4137 group.
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    Cell Signaling Technology Inc p gsk3β ser9
    A , B The activation level of <t>AKT/GSK3β</t> pathway was detected by western blot in vitro and in vivo. C , D The expression levels of EMT marker molecules (N-cadherin, E-cadherin and TGFβ1), inflammatory pathway protein molecules (NLRP3, Cleaved Caspase1/Caspase1) and fibrosis marker molecules (COL-1, COL-3 and α-SMA) was detected by western blot. E Inflammatory factor IL-1β and inflammatory chemokine CXCL2 the supernatant of alveolar epithelial cells were detected by ELISA. Data were expressed as the mean ± SD, n = 3. ** P < 0.01 and *** P < 0.001. F The migration and invasion abilities of alveolar epithelial cells were detected by scratch healing assay and transwell assay.
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    Cell Signaling Technology Inc p gsk3 β
    Protein expression in GA muscle samples from rats treated with n-3 PUFA or vehicle for 40 days and dexamethasone or vehicle (last 10 days). Relative optical density of total Akt (tAkt) and phosphorylated Akt (P-Akt) ((a) and (c)), total <t>GSK3</t> <t>β</t> (tGSK3 β ) and phosphorylated GSK3 β (P-GSK3 β ) ((b) and (d)), and tAkt/P-Akt and tGSK3 β /P-GSK3 β ratios ((e) and (f), resp.). Bands representing tAkt, P-Akt, tGSK3, and P-GSK3 β are illustrated in (g). α -Tubulin (55 kDa) was used as control. The numbers represent the mean ± S.E.M. CT = control group ( n = 6), DX = dexamethasone group ( n = 5), n-3 = n-3 PUFA group ( n = 6), and DX + n-3 = dexamethasone + n-3 PUFA group ( n = 6). Values represent means ± S.E.M., analyzed by Student's t -test, # P < 0.05.
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    Effects of DSG and 5-MOP on the expression of proteins in PI3K/Akt pathways involving ER α . Representative protein levels for (a) p-ER (Tyr-537)/ER α , (b) p-Src (Tyr-416)/Src, (c) PI3Kp85/ β -actin, (d) p-Akt (Ser473)/Akt, and (e) <t>p-GSK-3</t> β <t>(Ser9)/GSK-3</t> β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. ## P < 0.01: significance from DSG-treated group.
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    A, Western blots of MEK1/2, ERK1/2, p38, and JNK phosphorylation and their total protein levels 28 days after surgery in WT and KO mice. GAPDH was used as the sample loading control. (n = 6). Up, representative blots. Down, quantitative results. B, Western blots of <t>Akt,GSK3β,</t> and mTOR phosphorylation and their total protein levels 28 days after surgery. GAPDH was used as the sample loading control (n = 6). Up, representative blots. Down, quantitative results. For A and B, Data represent as the means ± SEM *P<0.01 vs. corresponding sham; #P<0.01 vs. WT/AB after AB.
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    Cell Signaling Technology Inc p gsk 3β ser9
    A and B, The phosphorylated and total protein levels of Akt, <t>GSK-3β,</t> mTOR, eIF4E, and 4E-BP1 in WT and Nek6 KO mice 4 weeks after TAC or sham surgery. Nek6 deficiency augments the activation of Akt signaling. A, representative western blots (duplicate lanes represent two different heart samples); B, quantitative results. * P <0.05 vs . WT/sham; # P <0.05 vs . WT/TAC. C and D, phosphorylated and total protein expression levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in H9c2 cells treated with Ang II for the indicated times with or without (control) the overexpression of Nek6. Nek6 overexpression attenuated the activation of Akt signaling. C, representative western blots; D, quantitative results. * P <0.05 vs. the control group at the 0 time point; # P <0.05 vs . the control group at the same time point.
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    Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, p-GSK3 β (Ser9), and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.

    Journal: Disease Markers

    Article Title: Leptin Promotes HTR-8/SVneo Cell Invasion via the Crosstalk between MTA1/WNT and PI3K/AKT Pathways

    doi: 10.1155/2022/7052176

    Figure Lengend Snippet: Leptin mediates β -catenin activation through the crosstalk between MTA1/WNT and PI3K/AKT pathways in HTR-8/SVneo cells. (a) HTR-8/SVneo cells were treated with exogenous leptin (0 and 200 ng/ml) for 24 h, and MTA1, WNT1, p-GSK3 β (Ser9), and p-AKT (Ser473) levels were detected by Western blot. Data are shown as mean ± SD; ∗ P < 0.01 vs. leptin (0 ng/ml). (b) The knockdown efficiencies of MTA1 and WNT1 were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (c) HTR-8/SVneo cells were transfected with MTA1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of MTA1, WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (d) HTR-8/SVneo cells were transfected with WNT1 siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of WNT1, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. (e) The knockdown efficiencies of AKT and β -catenin were analyzed by Western blot analysis and qRT-PCR. ∗ P < 0.05 vs. control. (f) HTR-8/SVneo cells were transfected with AKT siRNA or scramble siRNA (Scr) in the presence or absence of 200 ng/ml leptin for 24 h, and Western blot analysis was performed to detect the expression of AKT, p-GSK3 β (Ser9), and nuclear β -catenin. ∗ P < 0.01 vs. control and # P < 0.01 vs. leptin. All experiments were performed in triplicate.

    Article Snippet: The following primary antibodies were used: MMP9 (ab76003), MTA1 (ab71153), WNT1 (ab15251), AKT (ab179463), GSK3 β (ab32391), β -catenin (ab32572), histone 3 (ab1791), β -actin (ab6276) (Abcam, USA); p-AKT (Ser473) (#4060), p-GSK3 β (Ser9) (#9322) (Cell Signaling Technology, USA).

    Techniques: Activation Assay, Western Blot, Quantitative RT-PCR, Transfection, Expressing

    Effects of HA-ADT on the apoptotic level and AKT/GSK-3 β / β -catenin pathway in human HCC cells. (a) TUNEL staining was used to detect the apoptotic level (original magnification, 100x). (b) The apoptotic index was counted as the ratio of TUNEL positive cells to total cells. (c) Flow cytometry assay was adopted to detect apoptosis. (d) Cell apoptosis distribution was analyzed. (e) Western blotting was used to determine the protein levels of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, and p- β -catenin. β -actin was adopted as the internal control. (f) The density was analyzed. All data are shown as the mean ± SEM of three independent experiments; ∗ P < 0.05, ∗∗ P < 0.01 vs. control group; △ P < 0.05, △△ P < 0.01 vs. NaHS group; ## P < 0.01 vs. GYY4137 group.

    Journal: Journal of Oncology

    Article Title: A Water-Soluble Hydrogen Sulfide Donor Suppresses the Growth of Hepatocellular Carcinoma via Inhibiting the AKT/GSK-3 β / β -Catenin and TGF- β /Smad2/3 Signaling Pathways

    doi: 10.1155/2023/8456852

    Figure Lengend Snippet: Effects of HA-ADT on the apoptotic level and AKT/GSK-3 β / β -catenin pathway in human HCC cells. (a) TUNEL staining was used to detect the apoptotic level (original magnification, 100x). (b) The apoptotic index was counted as the ratio of TUNEL positive cells to total cells. (c) Flow cytometry assay was adopted to detect apoptosis. (d) Cell apoptosis distribution was analyzed. (e) Western blotting was used to determine the protein levels of AKT, p-AKT, GSK-3 β , p-GSK-3 β , β -catenin, and p- β -catenin. β -actin was adopted as the internal control. (f) The density was analyzed. All data are shown as the mean ± SEM of three independent experiments; ∗ P < 0.05, ∗∗ P < 0.01 vs. control group; △ P < 0.05, △△ P < 0.01 vs. NaHS group; ## P < 0.01 vs. GYY4137 group.

    Article Snippet: The primary antibodies include anti-Cyclin E1, anti-Cyclin D1, anti-cyclin-dependent kinase (CDK) 2, anti-CDK4, anti-p27, anti-p21, anti-AKT, anti-phospho (p)-AKT (Ser473), anti-glycogen synthase kinase-3 beta (Gsk‐3 β ), anti-p-Gsk-3 β (Ser9), anti- β -catenin, anti-p- β -catenin (Ser552), anti-beclin-1, anti-p62, anti-LC3A/B, anti-Smad2, anti-p-Smad2 (Ser465/467), anti-Smad3, anti-p-Smad3 (Ser423/425), and anti-transforming growth factor-beta (TGF‐ β ) antibodies, as well as the horseradish peroxidase-conjugated secondary antibody obtained from Cell Signaling Technology (CST, Danvers, MA, USA).

    Techniques: TUNEL Assay, Staining, Flow Cytometry, Western Blot

    A , B The activation level of AKT/GSK3β pathway was detected by western blot in vitro and in vivo. C , D The expression levels of EMT marker molecules (N-cadherin, E-cadherin and TGFβ1), inflammatory pathway protein molecules (NLRP3, Cleaved Caspase1/Caspase1) and fibrosis marker molecules (COL-1, COL-3 and α-SMA) was detected by western blot. E Inflammatory factor IL-1β and inflammatory chemokine CXCL2 the supernatant of alveolar epithelial cells were detected by ELISA. Data were expressed as the mean ± SD, n = 3. ** P < 0.01 and *** P < 0.001. F The migration and invasion abilities of alveolar epithelial cells were detected by scratch healing assay and transwell assay.

    Journal: Cell Death Discovery

    Article Title: Coal dust nanoparticles induced pulmonary fibrosis by promoting inflammation and epithelial-mesenchymal transition via the NF-κB/NLRP3 pathway driven by IGF1/ROS-mediated AKT/GSK3β signals

    doi: 10.1038/s41420-022-01291-z

    Figure Lengend Snippet: A , B The activation level of AKT/GSK3β pathway was detected by western blot in vitro and in vivo. C , D The expression levels of EMT marker molecules (N-cadherin, E-cadherin and TGFβ1), inflammatory pathway protein molecules (NLRP3, Cleaved Caspase1/Caspase1) and fibrosis marker molecules (COL-1, COL-3 and α-SMA) was detected by western blot. E Inflammatory factor IL-1β and inflammatory chemokine CXCL2 the supernatant of alveolar epithelial cells were detected by ELISA. Data were expressed as the mean ± SD, n = 3. ** P < 0.01 and *** P < 0.001. F The migration and invasion abilities of alveolar epithelial cells were detected by scratch healing assay and transwell assay.

    Article Snippet: The information of antibodies is as follows: IGF1 (1: 1000, Abclonal, #A11985, China), IGF1R (1: 1000, CST, #9750, USA), p-IGF1R(Tyr1135/1136) (1: 1000, CST, #3024, USA), COL-1 (1: 2000, Abcam, ab260043, UK), COL-3 (1: 1000, Abcam, ab184993, UK), α-SMA (1:1000, CST, #19425, USA), heme oxygenase-1 (HO-1, 1: 1000, CST, #5853, USA), N-cadherin (1: 1000, CST, #49398, USA), E-cadherin (1: 1000, CST, #49398, USA), TGFβ1 (1: 1000, abclonal, #A2124, China), p-NF-kB p65(Ser536) (1: 1000, CST, #3033, USA), NF-kB p65 (1: 1000, CST, #8242, USA), NLRP3 (1: 1000, CST, #15101, USA), Cleaved Caspase1(Asp297) (1: 1000, CST, #4199, USA), Caspase1 (1: 1000, CST, #2225, USA), p-Akt(Ser473) (1: 1000, CST, #4060, USA), Akt (1: 1000, CST, #4691, USA), p-GSK3β(Ser9) (1: 1000, CST, #9322, USA), GSK3β (1: 1000, CST, #9369, USA) and β-actin (1: 1000, CST, #4970, USA).

    Techniques: Activation Assay, Western Blot, In Vitro, In Vivo, Expressing, Marker, Enzyme-linked Immunosorbent Assay, Migration, Transwell Assay

    Protein expression in GA muscle samples from rats treated with n-3 PUFA or vehicle for 40 days and dexamethasone or vehicle (last 10 days). Relative optical density of total Akt (tAkt) and phosphorylated Akt (P-Akt) ((a) and (c)), total GSK3 β (tGSK3 β ) and phosphorylated GSK3 β (P-GSK3 β ) ((b) and (d)), and tAkt/P-Akt and tGSK3 β /P-GSK3 β ratios ((e) and (f), resp.). Bands representing tAkt, P-Akt, tGSK3, and P-GSK3 β are illustrated in (g). α -Tubulin (55 kDa) was used as control. The numbers represent the mean ± S.E.M. CT = control group ( n = 6), DX = dexamethasone group ( n = 5), n-3 = n-3 PUFA group ( n = 6), and DX + n-3 = dexamethasone + n-3 PUFA group ( n = 6). Values represent means ± S.E.M., analyzed by Student's t -test, # P < 0.05.

    Journal: BioMed Research International

    Article Title: The Effects of Omega-3 Fatty Acid Supplementation on Dexamethasone-Induced Muscle Atrophy

    doi: 10.1155/2014/961438

    Figure Lengend Snippet: Protein expression in GA muscle samples from rats treated with n-3 PUFA or vehicle for 40 days and dexamethasone or vehicle (last 10 days). Relative optical density of total Akt (tAkt) and phosphorylated Akt (P-Akt) ((a) and (c)), total GSK3 β (tGSK3 β ) and phosphorylated GSK3 β (P-GSK3 β ) ((b) and (d)), and tAkt/P-Akt and tGSK3 β /P-GSK3 β ratios ((e) and (f), resp.). Bands representing tAkt, P-Akt, tGSK3, and P-GSK3 β are illustrated in (g). α -Tubulin (55 kDa) was used as control. The numbers represent the mean ± S.E.M. CT = control group ( n = 6), DX = dexamethasone group ( n = 5), n-3 = n-3 PUFA group ( n = 6), and DX + n-3 = dexamethasone + n-3 PUFA group ( n = 6). Values represent means ± S.E.M., analyzed by Student's t -test, # P < 0.05.

    Article Snippet: The primary antibodies used were Akt (1 : 2,000, Cell Signaling #4691), P-Akt (1 : 2,000, Cell Signaling #4060, Ser473); mTOR (1 : 500, Cell Signaling #2972), P-mTOR (1 : 500, Cell Signaling #2971, Ser2448), GSK3 β (1 : 2,000, Cell Signaling #9315), P-GSK3 β (1 : 2,000, Cell Signaling #9322, Ser9), FoXO3a (1 : 500, Cell Signaling #2497), P-FoXO3a (1 : 500, Cell Signaling #9466, Ser253), and α -tubulin (1 : 30,000, Hybridoma bank) in 5% BSA/TBS-T.

    Techniques: Expressing

    Effects of DSG and 5-MOP on the expression of proteins in PI3K/Akt pathways involving ER α . Representative protein levels for (a) p-ER (Tyr-537)/ER α , (b) p-Src (Tyr-416)/Src, (c) PI3Kp85/ β -actin, (d) p-Akt (Ser473)/Akt, and (e) p-GSK-3 β (Ser9)/GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. ## P < 0.01: significance from DSG-treated group.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Diosgenin and 5-Methoxypsoralen Ameliorate Insulin Resistance through ER- α /PI3K/Akt-Signaling Pathways in HepG2 Cells

    doi: 10.1155/2016/7493694

    Figure Lengend Snippet: Effects of DSG and 5-MOP on the expression of proteins in PI3K/Akt pathways involving ER α . Representative protein levels for (a) p-ER (Tyr-537)/ER α , (b) p-Src (Tyr-416)/Src, (c) PI3Kp85/ β -actin, (d) p-Akt (Ser473)/Akt, and (e) p-GSK-3 β (Ser9)/GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. ## P < 0.01: significance from DSG-treated group.

    Article Snippet: Monoclonal antibody against Akt, p-Akt (Ser473), GSK-3 β , p-GSK-3 β (Ser9), Src, and p-Src (Tyr-416) were purchased from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Expressing

    Effects of DSG and 5-MOP on gene expression at the transcriptional level in PI3K/Akt pathways involving ER α . Representative mRNA levels for (a) ER α , (b) Src, (c) PI3K p85, (d) Akt, and (e) GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. # P < 0.05: significance from DSG-treated group.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Diosgenin and 5-Methoxypsoralen Ameliorate Insulin Resistance through ER- α /PI3K/Akt-Signaling Pathways in HepG2 Cells

    doi: 10.1155/2016/7493694

    Figure Lengend Snippet: Effects of DSG and 5-MOP on gene expression at the transcriptional level in PI3K/Akt pathways involving ER α . Representative mRNA levels for (a) ER α , (b) Src, (c) PI3K p85, (d) Akt, and (e) GSK-3 β . Each bar represents mean ± SD from three wells. ∗ P < 0.05 and ∗∗ P < 0.01: significance from control group. △ P < 0.05 and △△ P < 0.01: significance from model group. # P < 0.05: significance from DSG-treated group.

    Article Snippet: Monoclonal antibody against Akt, p-Akt (Ser473), GSK-3 β , p-GSK-3 β (Ser9), Src, and p-Src (Tyr-416) were purchased from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Expressing

    A, Western blots of MEK1/2, ERK1/2, p38, and JNK phosphorylation and their total protein levels 28 days after surgery in WT and KO mice. GAPDH was used as the sample loading control. (n = 6). Up, representative blots. Down, quantitative results. B, Western blots of Akt,GSK3β, and mTOR phosphorylation and their total protein levels 28 days after surgery. GAPDH was used as the sample loading control (n = 6). Up, representative blots. Down, quantitative results. For A and B, Data represent as the means ± SEM *P<0.01 vs. corresponding sham; #P<0.01 vs. WT/AB after AB.

    Journal: PLoS ONE

    Article Title: Augmented Cardiac Hypertrophy in Response to Pressure Overload in Mice Lacking ELTD1

    doi: 10.1371/journal.pone.0035779

    Figure Lengend Snippet: A, Western blots of MEK1/2, ERK1/2, p38, and JNK phosphorylation and their total protein levels 28 days after surgery in WT and KO mice. GAPDH was used as the sample loading control. (n = 6). Up, representative blots. Down, quantitative results. B, Western blots of Akt,GSK3β, and mTOR phosphorylation and their total protein levels 28 days after surgery. GAPDH was used as the sample loading control (n = 6). Up, representative blots. Down, quantitative results. For A and B, Data represent as the means ± SEM *P<0.01 vs. corresponding sham; #P<0.01 vs. WT/AB after AB.

    Article Snippet: Antibodies against the following proteins were purchased from Cell Signaling Technology: p-MEK1/2 (#9154), T-MEK1/2 (#9122), p-ERK1/2 (#4370), T-ERK1/2 (#4695), p-p38 (#4511), T-p38 (#9212), p-JNK1/2 (#4668), T-JNK 1/2(#9258), p-Akt (#4060), T-Akt (#4691), p-GSK3β (#9322), T-GSK3β (#9315), p-mTOR (#2974), T-mTOR (#2972), and GAPDH (#2118).

    Techniques: Western Blot

    A and B, The phosphorylated and total protein levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in WT and Nek6 KO mice 4 weeks after TAC or sham surgery. Nek6 deficiency augments the activation of Akt signaling. A, representative western blots (duplicate lanes represent two different heart samples); B, quantitative results. * P <0.05 vs . WT/sham; # P <0.05 vs . WT/TAC. C and D, phosphorylated and total protein expression levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in H9c2 cells treated with Ang II for the indicated times with or without (control) the overexpression of Nek6. Nek6 overexpression attenuated the activation of Akt signaling. C, representative western blots; D, quantitative results. * P <0.05 vs. the control group at the 0 time point; # P <0.05 vs . the control group at the same time point.

    Journal: PLoS ONE

    Article Title: Never in Mitosis Gene A Related Kinase-6 Attenuates Pressure Overload-Induced Activation of the Protein Kinase B Pathway and Cardiac Hypertrophy

    doi: 10.1371/journal.pone.0096095

    Figure Lengend Snippet: A and B, The phosphorylated and total protein levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in WT and Nek6 KO mice 4 weeks after TAC or sham surgery. Nek6 deficiency augments the activation of Akt signaling. A, representative western blots (duplicate lanes represent two different heart samples); B, quantitative results. * P <0.05 vs . WT/sham; # P <0.05 vs . WT/TAC. C and D, phosphorylated and total protein expression levels of Akt, GSK-3β, mTOR, eIF4E, and 4E-BP1 in H9c2 cells treated with Ang II for the indicated times with or without (control) the overexpression of Nek6. Nek6 overexpression attenuated the activation of Akt signaling. C, representative western blots; D, quantitative results. * P <0.05 vs. the control group at the 0 time point; # P <0.05 vs . the control group at the same time point.

    Article Snippet: The primary antibodies against phosphor (P)-Akt Thr308 (2965), total (T)-Akt (4691), P -GSK-3β Ser9 (9322), T-GSK-3β (9315), P-mTOR Ser2448 (2971), T-mTOR (2983), P-4EBP1 (2855p), T-4EBP1 (9644p), P-eIF4e (9741), T-eIF4e (2067) and GAPDH (2118) were purchased from Cell Signaling Technology.

    Techniques: Activation Assay, Western Blot, Expressing, Over Expression