phosphorylated as160 pas160 at s588  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc phosphorylated as160 pas160 at s588
    Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of <t>Insr/Akt/AS160</t> signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Phosphorylated As160 Pas160 At S588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated as160 pas160 at s588/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphorylated as160 pas160 at s588 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice"

    Article Title: Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice

    Journal: BMJ Open Diabetes Research & Care

    doi: 10.1136/bmjdrc-2020-001695

    Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of Insr/Akt/AS160 signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Figure Legend Snippet: Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of Insr/Akt/AS160 signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Techniques Used: Injection

    Activation of Insr and stimulation of Akt and AS160 phosphorylation in the skeletal muscle of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=3) or NPC43 (5.4 mpk, n=3) for 5 min. Gastrocnemius from these mice was collected and subjected to (A) western blot analysis by the indicated antibodies (using 75 µg protein/mouse). (B–D) Quantitative changes in protein expression of (B) pInsrβ-Y1146/Insrβ, (C) pAkt-T308/Akt, and (D) pAS160-S588/AS160 in the skeletal muscle of STZ-induced T1D mice after acute oral administration of NPC43, as determined by the western blot analysis shown in (A). Protein band densities in western blots were determined using National Institutes of Health (NIH) ImageJ software. For pAS160-S588 and AS160 protein expression, all protein bands detected by their specific monoclonal antibodies in western blots were quantitated. Data are presented as mean±SEM of three mice per group. P value (the NPC43-treated group vs the control saline group) was determined by performing Student’s t-test. Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Figure Legend Snippet: Activation of Insr and stimulation of Akt and AS160 phosphorylation in the skeletal muscle of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=3) or NPC43 (5.4 mpk, n=3) for 5 min. Gastrocnemius from these mice was collected and subjected to (A) western blot analysis by the indicated antibodies (using 75 µg protein/mouse). (B–D) Quantitative changes in protein expression of (B) pInsrβ-Y1146/Insrβ, (C) pAkt-T308/Akt, and (D) pAS160-S588/AS160 in the skeletal muscle of STZ-induced T1D mice after acute oral administration of NPC43, as determined by the western blot analysis shown in (A). Protein band densities in western blots were determined using National Institutes of Health (NIH) ImageJ software. For pAS160-S588 and AS160 protein expression, all protein bands detected by their specific monoclonal antibodies in western blots were quantitated. Data are presented as mean±SEM of three mice per group. P value (the NPC43-treated group vs the control saline group) was determined by performing Student’s t-test. Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Techniques Used: Activation Assay, Injection, Western Blot, Expressing, Software

    Activation of Insr and stimulation of Akt and AS160 phosphorylation in the liver of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=4) or NPC43 (5.4 mpk, n=4) for 3 hours. Blood glucose levels in these mice after saline and NPC43 treatments are shown in . Liver tissue from these mice was collected and subjected to western blot analysis of (A) activated Insr and (B) phosphorylated Akt and AS160 with the indicated antibodies (using 100 µg liver protein/mouse). Protein band densities in western blots shown in the left panels of A–B were determined using National Institutes of Health (NIH) ImageJ software, and data are presented as mean±SEM of four mice per group in the right bar graphs. In the bar graph shown in (B), ‘Ga.’ means Gapdh. *P<0.05, **P<0.01 versus the control saline group (Student’s t-test). Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Figure Legend Snippet: Activation of Insr and stimulation of Akt and AS160 phosphorylation in the liver of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=4) or NPC43 (5.4 mpk, n=4) for 3 hours. Blood glucose levels in these mice after saline and NPC43 treatments are shown in . Liver tissue from these mice was collected and subjected to western blot analysis of (A) activated Insr and (B) phosphorylated Akt and AS160 with the indicated antibodies (using 100 µg liver protein/mouse). Protein band densities in western blots shown in the left panels of A–B were determined using National Institutes of Health (NIH) ImageJ software, and data are presented as mean±SEM of four mice per group in the right bar graphs. In the bar graph shown in (B), ‘Ga.’ means Gapdh. *P<0.05, **P<0.01 versus the control saline group (Student’s t-test). Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Techniques Used: Activation Assay, Injection, Western Blot, Software

    phosphorylated as160 pas160 at s588  (Cell Signaling Technology Inc)


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    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Structured Review

    Cell Signaling Technology Inc phosphorylated as160 pas160 at s588
    Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of <t>Insr/Akt/AS160</t> signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Phosphorylated As160 Pas160 At S588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated as160 pas160 at s588/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphorylated as160 pas160 at s588 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice"

    Article Title: Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice

    Journal: BMJ Open Diabetes Research & Care

    doi: 10.1136/bmjdrc-2020-001695

    Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of Insr/Akt/AS160 signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Figure Legend Snippet: Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of Insr/Akt/AS160 signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Techniques Used: Injection

    Activation of Insr and stimulation of Akt and AS160 phosphorylation in the skeletal muscle of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=3) or NPC43 (5.4 mpk, n=3) for 5 min. Gastrocnemius from these mice was collected and subjected to (A) western blot analysis by the indicated antibodies (using 75 µg protein/mouse). (B–D) Quantitative changes in protein expression of (B) pInsrβ-Y1146/Insrβ, (C) pAkt-T308/Akt, and (D) pAS160-S588/AS160 in the skeletal muscle of STZ-induced T1D mice after acute oral administration of NPC43, as determined by the western blot analysis shown in (A). Protein band densities in western blots were determined using National Institutes of Health (NIH) ImageJ software. For pAS160-S588 and AS160 protein expression, all protein bands detected by their specific monoclonal antibodies in western blots were quantitated. Data are presented as mean±SEM of three mice per group. P value (the NPC43-treated group vs the control saline group) was determined by performing Student’s t-test. Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Figure Legend Snippet: Activation of Insr and stimulation of Akt and AS160 phosphorylation in the skeletal muscle of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=3) or NPC43 (5.4 mpk, n=3) for 5 min. Gastrocnemius from these mice was collected and subjected to (A) western blot analysis by the indicated antibodies (using 75 µg protein/mouse). (B–D) Quantitative changes in protein expression of (B) pInsrβ-Y1146/Insrβ, (C) pAkt-T308/Akt, and (D) pAS160-S588/AS160 in the skeletal muscle of STZ-induced T1D mice after acute oral administration of NPC43, as determined by the western blot analysis shown in (A). Protein band densities in western blots were determined using National Institutes of Health (NIH) ImageJ software. For pAS160-S588 and AS160 protein expression, all protein bands detected by their specific monoclonal antibodies in western blots were quantitated. Data are presented as mean±SEM of three mice per group. P value (the NPC43-treated group vs the control saline group) was determined by performing Student’s t-test. Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Techniques Used: Activation Assay, Injection, Western Blot, Expressing, Software

    Activation of Insr and stimulation of Akt and AS160 phosphorylation in the liver of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=4) or NPC43 (5.4 mpk, n=4) for 3 hours. Blood glucose levels in these mice after saline and NPC43 treatments are shown in . Liver tissue from these mice was collected and subjected to western blot analysis of (A) activated Insr and (B) phosphorylated Akt and AS160 with the indicated antibodies (using 100 µg liver protein/mouse). Protein band densities in western blots shown in the left panels of A–B were determined using National Institutes of Health (NIH) ImageJ software, and data are presented as mean±SEM of four mice per group in the right bar graphs. In the bar graph shown in (B), ‘Ga.’ means Gapdh. *P<0.05, **P<0.01 versus the control saline group (Student’s t-test). Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Figure Legend Snippet: Activation of Insr and stimulation of Akt and AS160 phosphorylation in the liver of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=4) or NPC43 (5.4 mpk, n=4) for 3 hours. Blood glucose levels in these mice after saline and NPC43 treatments are shown in . Liver tissue from these mice was collected and subjected to western blot analysis of (A) activated Insr and (B) phosphorylated Akt and AS160 with the indicated antibodies (using 100 µg liver protein/mouse). Protein band densities in western blots shown in the left panels of A–B were determined using National Institutes of Health (NIH) ImageJ software, and data are presented as mean±SEM of four mice per group in the right bar graphs. In the bar graph shown in (B), ‘Ga.’ means Gapdh. *P<0.05, **P<0.01 versus the control saline group (Student’s t-test). Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Techniques Used: Activation Assay, Injection, Western Blot, Software

    phospho as160  (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  
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    Structured Review

    Cell Signaling Technology Inc phospho as160
    Effect of Nar on HG-induced alterations of endothelial function. HUVECs were exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of Nar on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of Nar on intracellular MDA concentration. (d) Effect of Nar on NF- κ B activity. (e) Effect of Nar on IL-6 concentration in the culture media. (f) Effect of Nar on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of Nar on insulin-stimulated phosphorylation of Akt and its downstream <t>AS160.</t> (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of Nar on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus control (NG) group or the indicated group. ## P < 0.01 and ### P < 0.001 versus high-glucose (HG) group.
    Phospho As160, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho as160/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho as160 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Heat Shock Protein 70 Mediates the Protective Effect of Naringenin on High-Glucose-Induced Alterations of Endothelial Function"

    Article Title: Heat Shock Protein 70 Mediates the Protective Effect of Naringenin on High-Glucose-Induced Alterations of Endothelial Function

    Journal: International Journal of Endocrinology

    doi: 10.1155/2022/7275765

    Effect of Nar on HG-induced alterations of endothelial function. HUVECs were exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of Nar on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of Nar on intracellular MDA concentration. (d) Effect of Nar on NF- κ B activity. (e) Effect of Nar on IL-6 concentration in the culture media. (f) Effect of Nar on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of Nar on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of Nar on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus control (NG) group or the indicated group. ## P < 0.01 and ### P < 0.001 versus high-glucose (HG) group.
    Figure Legend Snippet: Effect of Nar on HG-induced alterations of endothelial function. HUVECs were exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of Nar on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of Nar on intracellular MDA concentration. (d) Effect of Nar on NF- κ B activity. (e) Effect of Nar on IL-6 concentration in the culture media. (f) Effect of Nar on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of Nar on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of Nar on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus control (NG) group or the indicated group. ## P < 0.01 and ### P < 0.001 versus high-glucose (HG) group.

    Techniques Used: Staining, Fluorescence, Concentration Assay, Activity Assay

    Effect of HSP70 knockdown (KD) on endothelial function. The siRNA technique was used to silence HSP70 in HUVECs. The cells were then grown in NG (5.5 mM glucose) media for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus siRNA control group or the indicated group.
    Figure Legend Snippet: Effect of HSP70 knockdown (KD) on endothelial function. The siRNA technique was used to silence HSP70 in HUVECs. The cells were then grown in NG (5.5 mM glucose) media for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus siRNA control group or the indicated group.

    Techniques Used: Staining, Fluorescence, Concentration Assay, Activity Assay

    HSP70 knockdown (KD) attenuated the protective effect of Nar on HG-induced alterations of endothelial function. The siRNA technique was used to silence HSP70 in HG-treated HUVECs. The cells were then exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus the indicated group.
    Figure Legend Snippet: HSP70 knockdown (KD) attenuated the protective effect of Nar on HG-induced alterations of endothelial function. The siRNA technique was used to silence HSP70 in HG-treated HUVECs. The cells were then exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus the indicated group.

    Techniques Used: Staining, Fluorescence, Concentration Assay, Activity Assay

    anti phospho as160 serine 588  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho as160 serine 588
    Effect of PHLPP1 silencing on PHLPP isoforms, Akt isoforms and <t>AS160</t> in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was silenced using PHLPP1 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in <t>pAS160</t> (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, # P < 0.05 as compared to lane 2, $$ P < 0.01, $$ P < 0.05 compared to lane 6. IB Immunoblot; IP Immunoprecipitation
    Anti Phospho As160 Serine 588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho as160 serine 588/product/Cell Signaling Technology Inc
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    1) Product Images from "PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble"

    Article Title: PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-022-00987-0

    Effect of PHLPP1 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was silenced using PHLPP1 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, # P < 0.05 as compared to lane 2, $$ P < 0.01, $$ P < 0.05 compared to lane 6. IB Immunoblot; IP Immunoprecipitation
    Figure Legend Snippet: Effect of PHLPP1 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was silenced using PHLPP1 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, # P < 0.05 as compared to lane 2, $$ P < 0.01, $$ P < 0.05 compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Techniques Used: Western Blot, Immunoprecipitation

    Effect of PHLPP2 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was silenced using PHLPP2 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$ P < 0.01 compared to lane 6 IB Immunoblot; IP Immunoprecipitation
    Figure Legend Snippet: Effect of PHLPP2 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was silenced using PHLPP2 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$ P < 0.01 compared to lane 6 IB Immunoblot; IP Immunoprecipitation

    Techniques Used: Western Blot, Immunoprecipitation

    Effect of PHLPP1 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was over-expressed using PHLPP1 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation
    Figure Legend Snippet: Effect of PHLPP1 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was over-expressed using PHLPP1 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Techniques Used: Over Expression, Plasmid Preparation, Western Blot, Immunoprecipitation

    Effect of PHLPP2 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was over-expressed using PHLPP2 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001, $$ P < 0.01 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation
    Figure Legend Snippet: Effect of PHLPP2 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was over-expressed using PHLPP2 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001, $$ P < 0.01 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Techniques Used: Over Expression, Plasmid Preparation, Western Blot, Immunoprecipitation

    Schematic diagram depicting role of PHLPP isoforms in regulating neuronal insulin signaling. A In an insulin sensitive, under unstimulated condition, PHLPP1 is localized on the membrane, while PHLPP2 is on the membrane as well as cytoplasm. Akt isoforms are present in the cytoplasm. AS160 binds to GSVs (GLUT4 Storage Vesicles) and tethers GLUT4. This does not allow GLUT4 exocytosis under basal conditions. B In an insulin resistant, unstimulated condition, hyperinsulinemia occurs due to defects in insulin signaling. PHLPP isoforms expression is elevated on the membrane. Scribble, the scaffolding protein holds PHLPP isoforms in place. C In an insulin sensitive, insulin stimulated condition, Akt translocates to plasma membrane in an isoform specific insulin dependent way (1), getting phosphorylated there (2). In neuronal system, all Akt isoforms translocate in the order Akt2 > Akt3 > Akt1. PHLPP1/2 are present on the membrane, constant flux of phosphorylation and dephosphorylation (3). An activated Akt phosphorylates AS160, hence inactivating it. Phosphorylated and thus inactivated AS160 translocated to cytoplasm (4), promoting GLUT4 exocytosis (5), and allowing glucose uptake (6). D In an insulin resistant, insulin stimulated condition, hyperinsulinemia triggers insulin receptor down-regulation, not allowing Akt isoform specific translocation to the plasma membrane (1). Elevated PHLPP isoforms dephosphorylate Akt isoforms remaining on the membrane (2). This leads to inadequate phosphorylation (3), leading to Akt’s inability to phosphorylate AS160 (4). Unphosphorylated, thus, active AS160 continues tethering GLUT4 to GSVs, not allowing GLUT4 exocytosis (5), hence affecting neuronal glucose uptake (6). (Created with BioRender.com)
    Figure Legend Snippet: Schematic diagram depicting role of PHLPP isoforms in regulating neuronal insulin signaling. A In an insulin sensitive, under unstimulated condition, PHLPP1 is localized on the membrane, while PHLPP2 is on the membrane as well as cytoplasm. Akt isoforms are present in the cytoplasm. AS160 binds to GSVs (GLUT4 Storage Vesicles) and tethers GLUT4. This does not allow GLUT4 exocytosis under basal conditions. B In an insulin resistant, unstimulated condition, hyperinsulinemia occurs due to defects in insulin signaling. PHLPP isoforms expression is elevated on the membrane. Scribble, the scaffolding protein holds PHLPP isoforms in place. C In an insulin sensitive, insulin stimulated condition, Akt translocates to plasma membrane in an isoform specific insulin dependent way (1), getting phosphorylated there (2). In neuronal system, all Akt isoforms translocate in the order Akt2 > Akt3 > Akt1. PHLPP1/2 are present on the membrane, constant flux of phosphorylation and dephosphorylation (3). An activated Akt phosphorylates AS160, hence inactivating it. Phosphorylated and thus inactivated AS160 translocated to cytoplasm (4), promoting GLUT4 exocytosis (5), and allowing glucose uptake (6). D In an insulin resistant, insulin stimulated condition, hyperinsulinemia triggers insulin receptor down-regulation, not allowing Akt isoform specific translocation to the plasma membrane (1). Elevated PHLPP isoforms dephosphorylate Akt isoforms remaining on the membrane (2). This leads to inadequate phosphorylation (3), leading to Akt’s inability to phosphorylate AS160 (4). Unphosphorylated, thus, active AS160 continues tethering GLUT4 to GSVs, not allowing GLUT4 exocytosis (5), hence affecting neuronal glucose uptake (6). (Created with BioRender.com)

    Techniques Used: Expressing, Scaffolding, De-Phosphorylation Assay, Translocation Assay

    anti phospho as160 ser 588  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho as160 ser 588
    (A) Phosphorylated <t>AS160</t> Ser704 /AS160; (B) Phosphorylated AS160 <t>Ser588</t> /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for <t>pAS160</t> Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.
    Anti Phospho As160 Ser 588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Prior AICAR induces elevated glucose uptake concomitant with greater γ3-AMPK activation and reduced membrane cholesterol in skeletal muscle from 26-month-old rats"

    Article Title: Prior AICAR induces elevated glucose uptake concomitant with greater γ3-AMPK activation and reduced membrane cholesterol in skeletal muscle from 26-month-old rats

    Journal: Facets (Ottawa)

    doi: 10.1139/facets-2021-0166

    (A) Phosphorylated AS160 Ser704 /AS160; (B) Phosphorylated AS160 Ser588 /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for pAS160 Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.
    Figure Legend Snippet: (A) Phosphorylated AS160 Ser704 /AS160; (B) Phosphorylated AS160 Ser588 /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for pAS160 Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.

    Techniques Used: Concentration Assay, Staining

    as160ser588 8730s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc as160ser588 8730s
    As160ser588 8730s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho as160 ser588  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho as160 ser588
    Phospho As160 Ser588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho akt substrate 160  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt substrate 160
    Phospho Akt Substrate 160, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ser588  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ser588
    Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) <t>AS160(Ser588),</t> and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.
    Ser588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Acute Effects of Cheddar Cheese Consumption on Circulating Amino Acids and Human Skeletal Muscle"

    Article Title: Acute Effects of Cheddar Cheese Consumption on Circulating Amino Acids and Human Skeletal Muscle

    Journal: Nutrients

    doi: 10.3390/nu13020614

    Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) AS160(Ser588), and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.
    Figure Legend Snippet: Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) AS160(Ser588), and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.

    Techniques Used: Expressing, Western Blot

    anti phospho as160 ser 588  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho as160 ser 588
    A–C: phosphorylated AS160Ser704, AS160Thr642, and <t>AS160Ser588/AS160</t> immediately postexercise. Data were analyzed by one-way ANOVA. *P < 0.005, HFD-IPEX vs. both LFD-SED and HFD-SED. †P < 0.005, HFD-IPEX vs. HFD-SED. Values are expressed as means ± SEM; n = 17/group. D–F: phosphorylated AS160Ser704, AS160Thr642, and AS160Ser588/AS160 at 3 h postexercise. Data were analyzed by two-way ANOVA. *P < 0.001, insulin vs. no insulin. †P < 0.001, HFD-SED vs. both LFD-SED and HFD-3hPEX within insulin-treated muscles. ‡P < 0.01, HFD-3hPEX vs. HFD-SED within insulin-treated muscles. Values are expressed as means ± SEM; n = 10–17/group. HFD, high-fat diet; LFD, low-fat diet; SED, sedentary; IPEX, immediately postexercise; 3hPEX, 3 h postexercise.
    Anti Phospho As160 Ser 588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Exercise effects on γ3-AMPK activity, phosphorylation of Akt2 and AS160, and insulin-stimulated glucose uptake in insulin-resistant rat skeletal muscle"

    Article Title: Exercise effects on γ3-AMPK activity, phosphorylation of Akt2 and AS160, and insulin-stimulated glucose uptake in insulin-resistant rat skeletal muscle

    Journal: Journal of Applied Physiology

    doi: 10.1152/japplphysiol.00428.2019

    A–C: phosphorylated AS160Ser704, AS160Thr642, and AS160Ser588/AS160 immediately postexercise. Data were analyzed by one-way ANOVA. *P < 0.005, HFD-IPEX vs. both LFD-SED and HFD-SED. †P < 0.005, HFD-IPEX vs. HFD-SED. Values are expressed as means ± SEM; n = 17/group. D–F: phosphorylated AS160Ser704, AS160Thr642, and AS160Ser588/AS160 at 3 h postexercise. Data were analyzed by two-way ANOVA. *P < 0.001, insulin vs. no insulin. †P < 0.001, HFD-SED vs. both LFD-SED and HFD-3hPEX within insulin-treated muscles. ‡P < 0.01, HFD-3hPEX vs. HFD-SED within insulin-treated muscles. Values are expressed as means ± SEM; n = 10–17/group. HFD, high-fat diet; LFD, low-fat diet; SED, sedentary; IPEX, immediately postexercise; 3hPEX, 3 h postexercise.
    Figure Legend Snippet: A–C: phosphorylated AS160Ser704, AS160Thr642, and AS160Ser588/AS160 immediately postexercise. Data were analyzed by one-way ANOVA. *P < 0.005, HFD-IPEX vs. both LFD-SED and HFD-SED. †P < 0.005, HFD-IPEX vs. HFD-SED. Values are expressed as means ± SEM; n = 17/group. D–F: phosphorylated AS160Ser704, AS160Thr642, and AS160Ser588/AS160 at 3 h postexercise. Data were analyzed by two-way ANOVA. *P < 0.001, insulin vs. no insulin. †P < 0.001, HFD-SED vs. both LFD-SED and HFD-3hPEX within insulin-treated muscles. ‡P < 0.01, HFD-3hPEX vs. HFD-SED within insulin-treated muscles. Values are expressed as means ± SEM; n = 10–17/group. HFD, high-fat diet; LFD, low-fat diet; SED, sedentary; IPEX, immediately postexercise; 3hPEX, 3 h postexercise.

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    anti tbc1d4 ser588 phosphorylation cell signaling technology  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tbc1d4 ser588 phosphorylation cell signaling technology
    Anti Tbc1d4 Ser588 Phosphorylation Cell Signaling Technology, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phosphorylated as160 pas160 at s588
    Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of <t>Insr/Akt/AS160</t> signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.
    Phosphorylated As160 Pas160 At S588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho as160
    Effect of Nar on HG-induced alterations of endothelial function. HUVECs were exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of Nar on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of Nar on intracellular MDA concentration. (d) Effect of Nar on NF- κ B activity. (e) Effect of Nar on IL-6 concentration in the culture media. (f) Effect of Nar on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of Nar on insulin-stimulated phosphorylation of Akt and its downstream <t>AS160.</t> (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of Nar on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus control (NG) group or the indicated group. ## P < 0.01 and ### P < 0.001 versus high-glucose (HG) group.
    Phospho As160, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho as160 serine 588
    Effect of PHLPP1 silencing on PHLPP isoforms, Akt isoforms and <t>AS160</t> in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was silenced using PHLPP1 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in <t>pAS160</t> (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, # P < 0.05 as compared to lane 2, $$ P < 0.01, $$ P < 0.05 compared to lane 6. IB Immunoblot; IP Immunoprecipitation
    Anti Phospho As160 Serine 588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho as160 ser 588
    (A) Phosphorylated <t>AS160</t> Ser704 /AS160; (B) Phosphorylated AS160 <t>Ser588</t> /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for <t>pAS160</t> Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.
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    Cell Signaling Technology Inc as160ser588 8730s
    (A) Phosphorylated <t>AS160</t> Ser704 /AS160; (B) Phosphorylated AS160 <t>Ser588</t> /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for <t>pAS160</t> Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.
    As160ser588 8730s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho as160 ser588
    (A) Phosphorylated <t>AS160</t> Ser704 /AS160; (B) Phosphorylated AS160 <t>Ser588</t> /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for <t>pAS160</t> Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.
    Phospho As160 Ser588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho akt substrate 160
    (A) Phosphorylated <t>AS160</t> Ser704 /AS160; (B) Phosphorylated AS160 <t>Ser588</t> /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for <t>pAS160</t> Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.
    Phospho Akt Substrate 160, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc ser588
    Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) <t>AS160(Ser588),</t> and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.
    Ser588, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti tbc1d4 ser588 phosphorylation cell signaling technology
    Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) <t>AS160(Ser588),</t> and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.
    Anti Tbc1d4 Ser588 Phosphorylation Cell Signaling Technology, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of Insr/Akt/AS160 signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Journal: BMJ Open Diabetes Research & Care

    Article Title: Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice

    doi: 10.1136/bmjdrc-2020-001695

    Figure Lengend Snippet: Reduced blood glucose levels in STZ-induced T1D mice after oral and intraperitoneal NPC43 treatments. (A, B) Change in blood glucose levels in STZ-induced mice after oral NPC43 administration. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At (A) 19–26 or (B) 48 days post-STZ treatment, STZ-induced T1D mice were fasted for 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline) or indicated dose(s) of NPC43. Blood glucose levels in STZ-induced T1D mice right before and after oral saline or NPC43 treatment at the indicated time points (under fasting conditions but having free access to water) were examined to obtain the change in blood glucose level in each mouse. Mean±SEM, *p<0.05, **p<0.01, ***p<0.001 versus the saline group at the same time point (Student’s t-test). The numbers in parenthesis are p values versus each treatment group before oral gavage (0 time point) (Student’s t-test). After blood glucose analysis in (B), liver tissues of STZ-induced T1D mice at 3 hours post saline or NPC43 treatments (four mice per group) were collected for protein analysis of Insr/Akt/AS160 signaling shown in . (C) Reduced blood glucose levels in STZ-induced T1D mice after intraperitoneal treatment with NPC43. STZ-induced T1D mice (at 14 days post-STZ treatment) with unfasted blood glucose levels between 500 and 550 mg/dL were fasted overnight and intraperitoneally injected with NPC43 (5.4 mpk) or 2% (v/v) DMSO/saline. At 1, 2 and 3 hours post-intraperitoneal NPC43 injection, blood glucose levels in these mice were measured using the glucometer. Data are presented as mean±SEM of four mice per group. P value (the NPC43-treated group vs the DMSO/saline group at the same time point) was determined by performing Student’s t-test. mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Article Snippet: Primary antibodies against phosphorylated Insrβ (pInsrβ) at Y1146 (Cat #3021), Insrβ (Cat #3025), phosphorylated Akt (pAkt) at T308 (Cat #2965), Akt (Cat #4691), phosphorylated AS160 (pAS160) at S588 (Cat #8730), AS160 (Cat #2670), and Gapdh (Cat #5174) were purchased from Cell Signaling Technology (Danvers, Massachusetts).

    Techniques: Injection

    Activation of Insr and stimulation of Akt and AS160 phosphorylation in the skeletal muscle of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=3) or NPC43 (5.4 mpk, n=3) for 5 min. Gastrocnemius from these mice was collected and subjected to (A) western blot analysis by the indicated antibodies (using 75 µg protein/mouse). (B–D) Quantitative changes in protein expression of (B) pInsrβ-Y1146/Insrβ, (C) pAkt-T308/Akt, and (D) pAS160-S588/AS160 in the skeletal muscle of STZ-induced T1D mice after acute oral administration of NPC43, as determined by the western blot analysis shown in (A). Protein band densities in western blots were determined using National Institutes of Health (NIH) ImageJ software. For pAS160-S588 and AS160 protein expression, all protein bands detected by their specific monoclonal antibodies in western blots were quantitated. Data are presented as mean±SEM of three mice per group. P value (the NPC43-treated group vs the control saline group) was determined by performing Student’s t-test. Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Journal: BMJ Open Diabetes Research & Care

    Article Title: Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice

    doi: 10.1136/bmjdrc-2020-001695

    Figure Lengend Snippet: Activation of Insr and stimulation of Akt and AS160 phosphorylation in the skeletal muscle of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=3) or NPC43 (5.4 mpk, n=3) for 5 min. Gastrocnemius from these mice was collected and subjected to (A) western blot analysis by the indicated antibodies (using 75 µg protein/mouse). (B–D) Quantitative changes in protein expression of (B) pInsrβ-Y1146/Insrβ, (C) pAkt-T308/Akt, and (D) pAS160-S588/AS160 in the skeletal muscle of STZ-induced T1D mice after acute oral administration of NPC43, as determined by the western blot analysis shown in (A). Protein band densities in western blots were determined using National Institutes of Health (NIH) ImageJ software. For pAS160-S588 and AS160 protein expression, all protein bands detected by their specific monoclonal antibodies in western blots were quantitated. Data are presented as mean±SEM of three mice per group. P value (the NPC43-treated group vs the control saline group) was determined by performing Student’s t-test. Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Article Snippet: Primary antibodies against phosphorylated Insrβ (pInsrβ) at Y1146 (Cat #3021), Insrβ (Cat #3025), phosphorylated Akt (pAkt) at T308 (Cat #2965), Akt (Cat #4691), phosphorylated AS160 (pAS160) at S588 (Cat #8730), AS160 (Cat #2670), and Gapdh (Cat #5174) were purchased from Cell Signaling Technology (Danvers, Massachusetts).

    Techniques: Activation Assay, Injection, Western Blot, Expressing, Software

    Activation of Insr and stimulation of Akt and AS160 phosphorylation in the liver of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=4) or NPC43 (5.4 mpk, n=4) for 3 hours. Blood glucose levels in these mice after saline and NPC43 treatments are shown in . Liver tissue from these mice was collected and subjected to western blot analysis of (A) activated Insr and (B) phosphorylated Akt and AS160 with the indicated antibodies (using 100 µg liver protein/mouse). Protein band densities in western blots shown in the left panels of A–B were determined using National Institutes of Health (NIH) ImageJ software, and data are presented as mean±SEM of four mice per group in the right bar graphs. In the bar graph shown in (B), ‘Ga.’ means Gapdh. *P<0.05, **P<0.01 versus the control saline group (Student’s t-test). Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Journal: BMJ Open Diabetes Research & Care

    Article Title: Oral administration of NPC43 counters hyperglycemia and activates insulin receptor in streptozotocin-induced type 1 diabetic mice

    doi: 10.1136/bmjdrc-2020-001695

    Figure Lengend Snippet: Activation of Insr and stimulation of Akt and AS160 phosphorylation in the liver of STZ-induced T1D mice by NPC43. Male C57BL6 mice (4–5 weeks old) were intraperitoneally injected with 55 mpk STZ daily for 5 days. At 48 days post-STZ treatment, animals were fasted 2 hours and then orally administered with 1% (v/v) dimethylsulfoxide (DMSO)/physiological saline (saline, n=4) or NPC43 (5.4 mpk, n=4) for 3 hours. Blood glucose levels in these mice after saline and NPC43 treatments are shown in . Liver tissue from these mice was collected and subjected to western blot analysis of (A) activated Insr and (B) phosphorylated Akt and AS160 with the indicated antibodies (using 100 µg liver protein/mouse). Protein band densities in western blots shown in the left panels of A–B were determined using National Institutes of Health (NIH) ImageJ software, and data are presented as mean±SEM of four mice per group in the right bar graphs. In the bar graph shown in (B), ‘Ga.’ means Gapdh. *P<0.05, **P<0.01 versus the control saline group (Student’s t-test). Insr, insulin receptor; mpk, mg/kg body weight; NPC43, adenosine, 5’-Se-methyl-5’-seleno-,2’,3’-diacetate; STZ, streptozotocin; T1D, type 1 diabetes.

    Article Snippet: Primary antibodies against phosphorylated Insrβ (pInsrβ) at Y1146 (Cat #3021), Insrβ (Cat #3025), phosphorylated Akt (pAkt) at T308 (Cat #2965), Akt (Cat #4691), phosphorylated AS160 (pAS160) at S588 (Cat #8730), AS160 (Cat #2670), and Gapdh (Cat #5174) were purchased from Cell Signaling Technology (Danvers, Massachusetts).

    Techniques: Activation Assay, Injection, Western Blot, Software

    Effect of Nar on HG-induced alterations of endothelial function. HUVECs were exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of Nar on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of Nar on intracellular MDA concentration. (d) Effect of Nar on NF- κ B activity. (e) Effect of Nar on IL-6 concentration in the culture media. (f) Effect of Nar on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of Nar on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of Nar on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus control (NG) group or the indicated group. ## P < 0.01 and ### P < 0.001 versus high-glucose (HG) group.

    Journal: International Journal of Endocrinology

    Article Title: Heat Shock Protein 70 Mediates the Protective Effect of Naringenin on High-Glucose-Induced Alterations of Endothelial Function

    doi: 10.1155/2022/7275765

    Figure Lengend Snippet: Effect of Nar on HG-induced alterations of endothelial function. HUVECs were exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of Nar on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of Nar on intracellular MDA concentration. (d) Effect of Nar on NF- κ B activity. (e) Effect of Nar on IL-6 concentration in the culture media. (f) Effect of Nar on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of Nar on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of Nar on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus control (NG) group or the indicated group. ## P < 0.01 and ### P < 0.001 versus high-glucose (HG) group.

    Article Snippet: Antibodies against Akt (#9272), AS160 (#2447), HSF1 (#4356), HSP70 (#4876), phospho-Akt (Thr308, #9275), and phospho-AS160 (Ser588, #8730) were obtained from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Staining, Fluorescence, Concentration Assay, Activity Assay

    Effect of HSP70 knockdown (KD) on endothelial function. The siRNA technique was used to silence HSP70 in HUVECs. The cells were then grown in NG (5.5 mM glucose) media for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus siRNA control group or the indicated group.

    Journal: International Journal of Endocrinology

    Article Title: Heat Shock Protein 70 Mediates the Protective Effect of Naringenin on High-Glucose-Induced Alterations of Endothelial Function

    doi: 10.1155/2022/7275765

    Figure Lengend Snippet: Effect of HSP70 knockdown (KD) on endothelial function. The siRNA technique was used to silence HSP70 in HUVECs. The cells were then grown in NG (5.5 mM glucose) media for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 versus siRNA control group or the indicated group.

    Article Snippet: Antibodies against Akt (#9272), AS160 (#2447), HSF1 (#4356), HSP70 (#4876), phospho-Akt (Thr308, #9275), and phospho-AS160 (Ser588, #8730) were obtained from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Staining, Fluorescence, Concentration Assay, Activity Assay

    HSP70 knockdown (KD) attenuated the protective effect of Nar on HG-induced alterations of endothelial function. The siRNA technique was used to silence HSP70 in HG-treated HUVECs. The cells were then exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus the indicated group.

    Journal: International Journal of Endocrinology

    Article Title: Heat Shock Protein 70 Mediates the Protective Effect of Naringenin on High-Glucose-Induced Alterations of Endothelial Function

    doi: 10.1155/2022/7275765

    Figure Lengend Snippet: HSP70 knockdown (KD) attenuated the protective effect of Nar on HG-induced alterations of endothelial function. The siRNA technique was used to silence HSP70 in HG-treated HUVECs. The cells were then exposed to NG (5.5 mM glucose) or HG (30 mM glucose) media with or without the 50 mM of Nar for 36 h. To observe the effect of Nar on insulin signaling, the cells were treated with or without 100 nM of insulin for 10 min. (a) Effect of HSP70 KD on ROS formation by DHE staining. (b) Quantification of relative DHE fluorescence in (a). (c) Effect of HSP70 KD on intracellular MDA concentration. (d) Effect of HSP70 KD on NF- κ B activity. (e) Effect of HSP70 KD on IL-6 concentration in the culture media. (f) Effect of HSP70 KD on mRNA levels of cell adhesion molecules ICAM-1 and VCAM-1. (g) Effect of HSP70 KD on insulin-stimulated phosphorylation of Akt and its downstream AS160. (h) Quantification of phosphorylated Akt and AS160 in (g). (i) Effect of HSP70 KD on insulin-stimulated 2-DG uptake. Data are expressed as means ± SD ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus the indicated group.

    Article Snippet: Antibodies against Akt (#9272), AS160 (#2447), HSF1 (#4356), HSP70 (#4876), phospho-Akt (Thr308, #9275), and phospho-AS160 (Ser588, #8730) were obtained from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Staining, Fluorescence, Concentration Assay, Activity Assay

    Effect of PHLPP1 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was silenced using PHLPP1 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, # P < 0.05 as compared to lane 2, $$ P < 0.01, $$ P < 0.05 compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Journal: Cell Communication and Signaling : CCS

    Article Title: PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble

    doi: 10.1186/s12964-022-00987-0

    Figure Lengend Snippet: Effect of PHLPP1 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was silenced using PHLPP1 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, # P < 0.05 as compared to lane 2, $$ P < 0.01, $$ P < 0.05 compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Article Snippet: Anti-phospho-Akt (serine-473) (Cat. No. 4058), anti-phospho-Akt1(Serine-473) (Cat. No. 9018), anti-phospho-Akt2 (Serine-474) (Cat. No. 8599), anti-Akt1 antibody (Cat. No. 2938), anti-Akt2 antibody (Cat. No. 3063), anti-phospho-AS160 (serine-588) (Cat. No. 8730), anti-phospho-AS160 (threonine-642) (Cat. No. 8881), anti-AS160 antibody (Cat. No. 2670), IgG conformational (Cat. No. 3678) were purchased from Cell Signalling Technology Inc. (USA).

    Techniques: Western Blot, Immunoprecipitation

    Effect of PHLPP2 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was silenced using PHLPP2 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$ P < 0.01 compared to lane 6 IB Immunoblot; IP Immunoprecipitation

    Journal: Cell Communication and Signaling : CCS

    Article Title: PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble

    doi: 10.1186/s12964-022-00987-0

    Figure Lengend Snippet: Effect of PHLPP2 silencing on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was silenced using PHLPP2 specific siRNA. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$ P < 0.01 compared to lane 6 IB Immunoblot; IP Immunoprecipitation

    Article Snippet: Anti-phospho-Akt (serine-473) (Cat. No. 4058), anti-phospho-Akt1(Serine-473) (Cat. No. 9018), anti-phospho-Akt2 (Serine-474) (Cat. No. 8599), anti-Akt1 antibody (Cat. No. 2938), anti-Akt2 antibody (Cat. No. 3063), anti-phospho-AS160 (serine-588) (Cat. No. 8730), anti-phospho-AS160 (threonine-642) (Cat. No. 8881), anti-AS160 antibody (Cat. No. 2670), IgG conformational (Cat. No. 3678) were purchased from Cell Signalling Technology Inc. (USA).

    Techniques: Western Blot, Immunoprecipitation

    Effect of PHLPP1 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was over-expressed using PHLPP1 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Journal: Cell Communication and Signaling : CCS

    Article Title: PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble

    doi: 10.1186/s12964-022-00987-0

    Figure Lengend Snippet: Effect of PHLPP1 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP1 was over-expressed using PHLPP1 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. B Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Article Snippet: Anti-phospho-Akt (serine-473) (Cat. No. 4058), anti-phospho-Akt1(Serine-473) (Cat. No. 9018), anti-phospho-Akt2 (Serine-474) (Cat. No. 8599), anti-Akt1 antibody (Cat. No. 2938), anti-Akt2 antibody (Cat. No. 3063), anti-phospho-AS160 (serine-588) (Cat. No. 8730), anti-phospho-AS160 (threonine-642) (Cat. No. 8881), anti-AS160 antibody (Cat. No. 2670), IgG conformational (Cat. No. 3678) were purchased from Cell Signalling Technology Inc. (USA).

    Techniques: Over Expression, Plasmid Preparation, Western Blot, Immunoprecipitation

    Effect of PHLPP2 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was over-expressed using PHLPP2 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001, $$ P < 0.01 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Journal: Cell Communication and Signaling : CCS

    Article Title: PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble

    doi: 10.1186/s12964-022-00987-0

    Figure Lengend Snippet: Effect of PHLPP2 over-expression on PHLPP isoforms, Akt isoforms and AS160 in insulin signaling and insulin resistant condition in neuronal cells (N2A). Three days post-proliferation, PHLPP2 was over-expressed using PHLPP2 specific plasmid. N2A cells were differentiated in serum-free medium in the absence of (MF) or chronic presence of 100 nM insulin (MFI) for 3 days. Cells were lysed and subjected to western blotting, followed by probing with relevant primary antibodies. A Bar represents relative change in PHLPP2 probed with anti-PHLPP2 antibody. B Bar represents relative change in PHLPP1 probed with anti-PHLPP1 antibody. C Bar represents relative change in pAkt1 (Ser-473) probed with anti-Akt1 antibody. D Bar represents relative change in pAkt2 (Ser-474) probed with anti-Akt2 antibody. E Post-insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in pAkt (Ser-473) probed with anti-Akt3 antibody. F Bar represents relative change in pAS160 (Ser-588) when probed with anti-AS160 antibody. G Bar represents relative change in pAS160 (Thr-642) when probed with anti-AS160 antibody. GAPDH has been used as a loading control. Experiments were executed three times and a representative result is shown. Data expressed are mean ± SE. *** P < 0.001, ** P < 0.01, * P < 0.05 compared to lane 1, ### P < 0.001, ## P < 0.01, # P < 0.05 as compared to lane 2, $$$ P < 0.001, $$ P < 0.01 as compared to lane 6. IB Immunoblot; IP Immunoprecipitation

    Article Snippet: Anti-phospho-Akt (serine-473) (Cat. No. 4058), anti-phospho-Akt1(Serine-473) (Cat. No. 9018), anti-phospho-Akt2 (Serine-474) (Cat. No. 8599), anti-Akt1 antibody (Cat. No. 2938), anti-Akt2 antibody (Cat. No. 3063), anti-phospho-AS160 (serine-588) (Cat. No. 8730), anti-phospho-AS160 (threonine-642) (Cat. No. 8881), anti-AS160 antibody (Cat. No. 2670), IgG conformational (Cat. No. 3678) were purchased from Cell Signalling Technology Inc. (USA).

    Techniques: Over Expression, Plasmid Preparation, Western Blot, Immunoprecipitation

    Schematic diagram depicting role of PHLPP isoforms in regulating neuronal insulin signaling. A In an insulin sensitive, under unstimulated condition, PHLPP1 is localized on the membrane, while PHLPP2 is on the membrane as well as cytoplasm. Akt isoforms are present in the cytoplasm. AS160 binds to GSVs (GLUT4 Storage Vesicles) and tethers GLUT4. This does not allow GLUT4 exocytosis under basal conditions. B In an insulin resistant, unstimulated condition, hyperinsulinemia occurs due to defects in insulin signaling. PHLPP isoforms expression is elevated on the membrane. Scribble, the scaffolding protein holds PHLPP isoforms in place. C In an insulin sensitive, insulin stimulated condition, Akt translocates to plasma membrane in an isoform specific insulin dependent way (1), getting phosphorylated there (2). In neuronal system, all Akt isoforms translocate in the order Akt2 > Akt3 > Akt1. PHLPP1/2 are present on the membrane, constant flux of phosphorylation and dephosphorylation (3). An activated Akt phosphorylates AS160, hence inactivating it. Phosphorylated and thus inactivated AS160 translocated to cytoplasm (4), promoting GLUT4 exocytosis (5), and allowing glucose uptake (6). D In an insulin resistant, insulin stimulated condition, hyperinsulinemia triggers insulin receptor down-regulation, not allowing Akt isoform specific translocation to the plasma membrane (1). Elevated PHLPP isoforms dephosphorylate Akt isoforms remaining on the membrane (2). This leads to inadequate phosphorylation (3), leading to Akt’s inability to phosphorylate AS160 (4). Unphosphorylated, thus, active AS160 continues tethering GLUT4 to GSVs, not allowing GLUT4 exocytosis (5), hence affecting neuronal glucose uptake (6). (Created with BioRender.com)

    Journal: Cell Communication and Signaling : CCS

    Article Title: PHLPP isoforms differentially regulate Akt isoforms and AS160 affecting neuronal insulin signaling and insulin resistance via Scribble

    doi: 10.1186/s12964-022-00987-0

    Figure Lengend Snippet: Schematic diagram depicting role of PHLPP isoforms in regulating neuronal insulin signaling. A In an insulin sensitive, under unstimulated condition, PHLPP1 is localized on the membrane, while PHLPP2 is on the membrane as well as cytoplasm. Akt isoforms are present in the cytoplasm. AS160 binds to GSVs (GLUT4 Storage Vesicles) and tethers GLUT4. This does not allow GLUT4 exocytosis under basal conditions. B In an insulin resistant, unstimulated condition, hyperinsulinemia occurs due to defects in insulin signaling. PHLPP isoforms expression is elevated on the membrane. Scribble, the scaffolding protein holds PHLPP isoforms in place. C In an insulin sensitive, insulin stimulated condition, Akt translocates to plasma membrane in an isoform specific insulin dependent way (1), getting phosphorylated there (2). In neuronal system, all Akt isoforms translocate in the order Akt2 > Akt3 > Akt1. PHLPP1/2 are present on the membrane, constant flux of phosphorylation and dephosphorylation (3). An activated Akt phosphorylates AS160, hence inactivating it. Phosphorylated and thus inactivated AS160 translocated to cytoplasm (4), promoting GLUT4 exocytosis (5), and allowing glucose uptake (6). D In an insulin resistant, insulin stimulated condition, hyperinsulinemia triggers insulin receptor down-regulation, not allowing Akt isoform specific translocation to the plasma membrane (1). Elevated PHLPP isoforms dephosphorylate Akt isoforms remaining on the membrane (2). This leads to inadequate phosphorylation (3), leading to Akt’s inability to phosphorylate AS160 (4). Unphosphorylated, thus, active AS160 continues tethering GLUT4 to GSVs, not allowing GLUT4 exocytosis (5), hence affecting neuronal glucose uptake (6). (Created with BioRender.com)

    Article Snippet: Anti-phospho-Akt (serine-473) (Cat. No. 4058), anti-phospho-Akt1(Serine-473) (Cat. No. 9018), anti-phospho-Akt2 (Serine-474) (Cat. No. 8599), anti-Akt1 antibody (Cat. No. 2938), anti-Akt2 antibody (Cat. No. 3063), anti-phospho-AS160 (serine-588) (Cat. No. 8730), anti-phospho-AS160 (threonine-642) (Cat. No. 8881), anti-AS160 antibody (Cat. No. 2670), IgG conformational (Cat. No. 3678) were purchased from Cell Signalling Technology Inc. (USA).

    Techniques: Expressing, Scaffolding, De-Phosphorylation Assay, Translocation Assay

    (A) Phosphorylated AS160 Ser704 /AS160; (B) Phosphorylated AS160 Ser588 /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for pAS160 Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.

    Journal: Facets (Ottawa)

    Article Title: Prior AICAR induces elevated glucose uptake concomitant with greater γ3-AMPK activation and reduced membrane cholesterol in skeletal muscle from 26-month-old rats

    doi: 10.1139/facets-2021-0166

    Figure Lengend Snippet: (A) Phosphorylated AS160 Ser704 /AS160; (B) Phosphorylated AS160 Ser588 /AS160 and (C) Phosphorylated AS160 Thr642 /AS160, in epitrochlearis muscles. *Insulin versus without insulin within the same 5-aminoimidazole-4-carboxamide-1- β -d-ribofuranoside (AICAR) treatment ( P < 0.01 with AICAR for pAS160 Ser704 /AS160; P < 0.01 for pAS160 Ser588 /AS160, P < 0.001 for pAS160 Thr642 /AS160). † AICAR versus without AICAR with the same insulin concentration ( P < 0.05 without insulin and P < 0.001 with insulin for pAS160 Ser704 /AS160; P < 0.01 with insulin for pAS160 Thr642 /AS160). Data were analyzed using two-way analysis (insulin × AICAR) of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 7 per treatment group. The figure includes representative blots of phosphorylated signaling proteins and corresponding total signaling proteins below the graph. Total protein (based on MemCode staining) served as the loading control. Note: Akt substrate of 160 kDa, AS160.

    Article Snippet: Anti-phospho Akt Ser 473 (pAkt Ser473 ; #9271), anti-phospho Akt Thr 308 (pAkt Thr308 ; #13038), anti-Akt (#4691), anti-phospho AS160 Thr 642 (pAS160 Thr642 ; #8881), anti-phospho AS160 Ser 588 (pAS160 Ser588 ; #8730), anti-phospho AMPKα Thr 172 (pAMPKa Thr172 ; #2531), anti-AMP-activated protein kinase-α (AMPKα; #5831), anti-acetyl CoA carboxylase (ACC; #3676), anti-phospho ACC Ser79 (pACC Ser79 ; #3661), anti-TBC1D1 (#5929), anti-hexokinase II (HKII; #2867), anti-insulin receptor (IR; #3025), anti-α-Tubulin (#2144), anti-Na + /K + -ATPase (#3010) and anti-rabbit IgG horseradish peroxidase conjugate (#7074) were from Cell Signaling Technology (Danvers, MA).

    Techniques: Concentration Assay, Staining

    Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) AS160(Ser588), and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.

    Journal: Nutrients

    Article Title: Acute Effects of Cheddar Cheese Consumption on Circulating Amino Acids and Human Skeletal Muscle

    doi: 10.3390/nu13020614

    Figure Lengend Snippet: Skeletal muscle protein expression using immunoblotting for ( A ) p70S6K1(Thr389), ( B ) rpS6(Ser240/244), ( C ) Akt(Ser473), ( D ) AS160(Ser588), and ( E ) GSK-3β(Ser9) in the fasted state (0 min) and at 60 and 180 min following the ingestion of either cheese (solid line) or milk (dotted line) in men and women. Panels ( A , B ) are data for n = 24 while for ( C – E ) only n = 8 (4 M, 4 F) were analyzed. Panel ( F ) are representative images of immunoblotting. Phosphorylated protein levels were normalized to Ponceau-S. Different from baseline (0 min) for milk (*), p < 0.05. # , Different between groups at the specific time point, p < 0.05.

    Article Snippet: The primary antibodies were purchased from Cell Signaling Technology and were the following: phospho-S6K1, Thr389, 1:1000, #9205; phospho-ribosomal protein S6, RPS6, Ser240/244, 1:1000, #2215; phospho-AS160, Ser588, 1:1000, #8730; phospho-GSK-3β, Ser9, 1:1000, #9336; phospho-Akt, Ser473, 1:1000, #9271.

    Techniques: Expressing, Western Blot