total akt2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc total akt2
    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, <t>p-Akt2,</t> and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
    Total Akt2, 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/total akt2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    total akt2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice"

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.12101

    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
    Figure Legend Snippet: Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Techniques Used: Expressing, Western Blot

    Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.
    Figure Legend Snippet: Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Techniques Used: Expressing, Western Blot

    A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.
    Figure Legend Snippet: A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Techniques Used: Activity Assay, Expressing

    total akt2  (Cell Signaling Technology Inc)


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  • 94

    Structured Review

    Cell Signaling Technology Inc total akt2
    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, <t>p-Akt2,</t> and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
    Total Akt2, 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/total akt2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    total akt2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice"

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.12101

    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
    Figure Legend Snippet: Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Techniques Used: Expressing, Western Blot

    Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.
    Figure Legend Snippet: Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Techniques Used: Expressing, Western Blot

    A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.
    Figure Legend Snippet: A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Techniques Used: Activity Assay, Expressing

    phospho akt2  (Cell Signaling Technology Inc)


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  • 94

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    Cell Signaling Technology Inc phospho akt2
    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, <t>p-Akt2,</t> and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
    Phospho Akt2, 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 akt2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice"

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.12101

    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
    Figure Legend Snippet: Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Techniques Used: Expressing, Western Blot

    Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.
    Figure Legend Snippet: Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Techniques Used: Expressing, Western Blot

    A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.
    Figure Legend Snippet: A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Techniques Used: Activity Assay, Expressing

    ser 474  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ser 474
    Ser 474, 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/ser 474/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ser 474 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    phospho akt2 s473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt2 s473
    KEY RESOURCES TABLE
    Phospho Akt2 S473, 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 akt2 s473/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt2 s473 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake"

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.110509

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.
    Figure Legend Snippet: (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Techniques Used: Western Blot

    (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.
    Figure Legend Snippet: (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Techniques Used:

    (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).
    Figure Legend Snippet: (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Techniques Used:

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    phospho akt2 t308  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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  • 94

    Structured Review

    Cell Signaling Technology Inc phospho akt2 t308
    KEY RESOURCES TABLE
    Phospho Akt2 T308, 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 akt2 t308/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt2 t308 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake"

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.110509

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.
    Figure Legend Snippet: (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Techniques Used: Western Blot

    (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.
    Figure Legend Snippet: (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Techniques Used:

    (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).
    Figure Legend Snippet: (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Techniques Used:

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    phospho akt2 s473  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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  • 94

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    Cell Signaling Technology Inc phospho akt2 s473
    KEY RESOURCES TABLE
    Phospho Akt2 S473, 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 akt2 s473/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt2 s473 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake"

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.110509

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.
    Figure Legend Snippet: (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Techniques Used: Western Blot

    (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.
    Figure Legend Snippet: (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Techniques Used:

    (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).
    Figure Legend Snippet: (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Techniques Used:

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    phospho akt2 t308  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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  • 94

    Structured Review

    Cell Signaling Technology Inc phospho akt2 t308
    KEY RESOURCES TABLE
    Phospho Akt2 T308, 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 akt2 t308/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho akt2 t308 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake"

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.110509

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.
    Figure Legend Snippet: (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Techniques Used: Western Blot

    (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.
    Figure Legend Snippet: (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Techniques Used:

    (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).
    Figure Legend Snippet: (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Techniques Used:

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    p akt2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p akt2
    Composition of Akt isoforms in primary human subcutaneous preadipocytes. ( A – D ) Electropherograms of capillary isoelectric focusing (cIEF) immunoassays of ( A ) Akt1, ( B ) <t>Akt2,</t> ( C ) Akt3, and ( D ) pan-Akt or all Akt isoforms. The letter p represents phosphorylation of Akt isoforms. Peaks on electropherograms are color-coded with Akt1 (blue), Akt2 (green), and Akt3 (red). ( E ) Percentage of Akt1 (blue), Akt2 (green), and Akt3 (red) as a function of all Akt isoforms. ( F – H ) Electropherograms of cIEF immunoassays of ( F ) pAkt (Thr308), ( G ) pAkt (Thr450), and ( H ) pAkt (S473). The dashed line highlights the distribution of pAkt (Thr450) as a function of pI values.
    P Akt2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p akt2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p akt2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation"

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    Journal: Nutrients

    doi: 10.3390/nu14163301

    Composition of Akt isoforms in primary human subcutaneous preadipocytes. ( A – D ) Electropherograms of capillary isoelectric focusing (cIEF) immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, and ( D ) pan-Akt or all Akt isoforms. The letter p represents phosphorylation of Akt isoforms. Peaks on electropherograms are color-coded with Akt1 (blue), Akt2 (green), and Akt3 (red). ( E ) Percentage of Akt1 (blue), Akt2 (green), and Akt3 (red) as a function of all Akt isoforms. ( F – H ) Electropherograms of cIEF immunoassays of ( F ) pAkt (Thr308), ( G ) pAkt (Thr450), and ( H ) pAkt (S473). The dashed line highlights the distribution of pAkt (Thr450) as a function of pI values.
    Figure Legend Snippet: Composition of Akt isoforms in primary human subcutaneous preadipocytes. ( A – D ) Electropherograms of capillary isoelectric focusing (cIEF) immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, and ( D ) pan-Akt or all Akt isoforms. The letter p represents phosphorylation of Akt isoforms. Peaks on electropherograms are color-coded with Akt1 (blue), Akt2 (green), and Akt3 (red). ( E ) Percentage of Akt1 (blue), Akt2 (green), and Akt3 (red) as a function of all Akt isoforms. ( F – H ) Electropherograms of cIEF immunoassays of ( F ) pAkt (Thr308), ( G ) pAkt (Thr450), and ( H ) pAkt (S473). The dashed line highlights the distribution of pAkt (Thr450) as a function of pI values.

    Techniques Used:

    Insulin induces changes to the electropherograms of Akt2 and Akt3. Electropherograms of cIEF immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, ( D ) p-Akt (Thr308), ( E ) p-Akt (Thr450), and ( F ) p-Akt (Ser473) following treatment with insulin for 30 min. Arrows point to new peaks that appeared after treatment with insulin. The dashed line highlights the distribution of p-Akt (Thr450) as a function of pI values. ( G ) Capillary Western (CW) immunoassays of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before and after treatment with insulin. Pan-Akt and β-actin served as the loading controls. ( H ) Relative expression level of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before (solid black) and after (textured) treatment with insulin. Error bars indicate standard deviations across six repeated measurements using CW immunoassays per experimental condition. Asterisks indicate a statistical significance of p ≤ 0.01 versus untreated control.
    Figure Legend Snippet: Insulin induces changes to the electropherograms of Akt2 and Akt3. Electropherograms of cIEF immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, ( D ) p-Akt (Thr308), ( E ) p-Akt (Thr450), and ( F ) p-Akt (Ser473) following treatment with insulin for 30 min. Arrows point to new peaks that appeared after treatment with insulin. The dashed line highlights the distribution of p-Akt (Thr450) as a function of pI values. ( G ) Capillary Western (CW) immunoassays of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before and after treatment with insulin. Pan-Akt and β-actin served as the loading controls. ( H ) Relative expression level of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before (solid black) and after (textured) treatment with insulin. Error bars indicate standard deviations across six repeated measurements using CW immunoassays per experimental condition. Asterisks indicate a statistical significance of p ≤ 0.01 versus untreated control.

    Techniques Used: Western Blot, Expressing

    Cinnamaldehyde and curcumin induce changes to Akt2 posttranslational modification. ( A – E ) Distribution of Akt2 as a function of isoelectric points in ( A ) control untreated preadipocytes, or ( B – E ) preadipocytes treated with ( B ) insulin, ( C ) cinnamaldehyde, ( D ) curcumin, or ( E ) combined cinnamaldehyde and curcumin. ( F ) Distribution of p-Akt2 (S474) as a function of isoelectric points in untreated control preadipocytes (top electropherogram), or preadipocytes treated with insulin (second electropherogram), cinnamaldehyde (third electropherogram), curcumin (fourth electropherogram), and combined cinnamaldehyde and curcumin (bottom electropherogram). Arrows point to the appearance of new peaks following treatment versus untreated control.
    Figure Legend Snippet: Cinnamaldehyde and curcumin induce changes to Akt2 posttranslational modification. ( A – E ) Distribution of Akt2 as a function of isoelectric points in ( A ) control untreated preadipocytes, or ( B – E ) preadipocytes treated with ( B ) insulin, ( C ) cinnamaldehyde, ( D ) curcumin, or ( E ) combined cinnamaldehyde and curcumin. ( F ) Distribution of p-Akt2 (S474) as a function of isoelectric points in untreated control preadipocytes (top electropherogram), or preadipocytes treated with insulin (second electropherogram), cinnamaldehyde (third electropherogram), curcumin (fourth electropherogram), and combined cinnamaldehyde and curcumin (bottom electropherogram). Arrows point to the appearance of new peaks following treatment versus untreated control.

    Techniques Used: Modification

    Cinnamaldehyde and curcumin enhance insulin-stimulated activation of Akt2. ( A – D ) Distribution of Akt2 as a function of isoelectric points in preadipocytes treated with ( A ) insulin alone, ( B ) insulin and cinnamaldehyde, ( C ) insulin and curcumin, or ( D ) insulin, cinnamaldehyde and curcumin. ( E ) Distribution of p-Akt2 (S474) as a function of isoelectric points in preadipocytes treated with insulin alone (top electropherogram), insulin and cinnamaldehyde (second electropherogram), insulin and curcumin (third electropherogram), or insulin, cinnamaldehyde, and curcumin (bottom electropherogram). ( F ) Relative abundance of p-Akt2 (S474) as a function of treatment condition. Error bars are standard deviations across nine repeated measurements. Single asterisk (*) indicates p -value ≤ 0.01 versus treatment with insulin alone. Double asterisk (**) indicates p -value ≤ 0.01 versus treatment with insulin and cinnamaldehyde or insulin and curcumin.
    Figure Legend Snippet: Cinnamaldehyde and curcumin enhance insulin-stimulated activation of Akt2. ( A – D ) Distribution of Akt2 as a function of isoelectric points in preadipocytes treated with ( A ) insulin alone, ( B ) insulin and cinnamaldehyde, ( C ) insulin and curcumin, or ( D ) insulin, cinnamaldehyde and curcumin. ( E ) Distribution of p-Akt2 (S474) as a function of isoelectric points in preadipocytes treated with insulin alone (top electropherogram), insulin and cinnamaldehyde (second electropherogram), insulin and curcumin (third electropherogram), or insulin, cinnamaldehyde, and curcumin (bottom electropherogram). ( F ) Relative abundance of p-Akt2 (S474) as a function of treatment condition. Error bars are standard deviations across nine repeated measurements. Single asterisk (*) indicates p -value ≤ 0.01 versus treatment with insulin alone. Double asterisk (**) indicates p -value ≤ 0.01 versus treatment with insulin and cinnamaldehyde or insulin and curcumin.

    Techniques Used: Activation Assay

    Cinnamaldehyde and curcumin increase pan-Akt phosphorylation. ( A – C ) Distribution of p-Akt (T450) as a function of isoelectric points in ( A ) untreated control preadipocytes or ( B , C ) preadipocytes treated with ( B ) cinnamaldehyde or ( C ) curcumin. The dashed line highlights the distribution trend. ( D – F ) Distribution of ( D ) p-Akt (S124), ( E ) p-Akt (S246), and ( F ) p-Akt (Y475) in untreated control preadipocytes (top electropherogram), or preadipocytes treated with cinnamaldehyde (middle electropherogram) or curcumin (bottom electropherogram).
    Figure Legend Snippet: Cinnamaldehyde and curcumin increase pan-Akt phosphorylation. ( A – C ) Distribution of p-Akt (T450) as a function of isoelectric points in ( A ) untreated control preadipocytes or ( B , C ) preadipocytes treated with ( B ) cinnamaldehyde or ( C ) curcumin. The dashed line highlights the distribution trend. ( D – F ) Distribution of ( D ) p-Akt (S124), ( E ) p-Akt (S246), and ( F ) p-Akt (Y475) in untreated control preadipocytes (top electropherogram), or preadipocytes treated with cinnamaldehyde (middle electropherogram) or curcumin (bottom electropherogram).

    Techniques Used:

    Dual inhibition of PP2A and PTP1B activates Akt2. ( A – C ) Distribution of Akt2 as a function of isoelectric points in control untreated preadipocytes (top electropherograms) or in preadipocytes treated with ( A ) okadaic acid, ( B ) PTP1Bi, or ( C ) combined okadaic acid and PTP1B (bottom electropherograms). Arrows point to new peaks or peaks that experience elevated abundance in treated preadipocytes versus untreated control preadipocytes. ( D ) Distribution of p-Akt2 (S474) as a function of isoelectric point in control untreated preadipocytes (top electropherogram), or preadipocytes treated with okadaic acid (second electropherogram), PTP1Bi (third electropherogram), or combined okadaic acid and PTP1Bi (bottom electropherogram).
    Figure Legend Snippet: Dual inhibition of PP2A and PTP1B activates Akt2. ( A – C ) Distribution of Akt2 as a function of isoelectric points in control untreated preadipocytes (top electropherograms) or in preadipocytes treated with ( A ) okadaic acid, ( B ) PTP1Bi, or ( C ) combined okadaic acid and PTP1B (bottom electropherograms). Arrows point to new peaks or peaks that experience elevated abundance in treated preadipocytes versus untreated control preadipocytes. ( D ) Distribution of p-Akt2 (S474) as a function of isoelectric point in control untreated preadipocytes (top electropherogram), or preadipocytes treated with okadaic acid (second electropherogram), PTP1Bi (third electropherogram), or combined okadaic acid and PTP1Bi (bottom electropherogram).

    Techniques Used: Inhibition

    pathscan phospho akt2 ser474  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pathscan phospho akt2 ser474
    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of <t>AKT2</t> normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.
    Pathscan Phospho Akt2 Ser474, 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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    Images

    1) Product Images from "Development of a physiological insulin resistance model in human stem cell–derived adipocytes"

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    Journal: Science Advances

    doi: 10.1126/sciadv.abn7298

    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.
    Figure Legend Snippet: ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.

    Techniques Used: Functional Assay

    ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.
    Figure Legend Snippet: ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.

    Techniques Used: Translocation Assay, Two Tailed Test

    ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.
    Figure Legend Snippet: ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.

    Techniques Used:

    akt2 levels  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc akt2 levels
    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of <t>AKT2</t> normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.
    Akt2 Levels, 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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    Images

    1) Product Images from "Development of a physiological insulin resistance model in human stem cell–derived adipocytes"

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    Journal: Science Advances

    doi: 10.1126/sciadv.abn7298

    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.
    Figure Legend Snippet: ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.

    Techniques Used: Functional Assay

    ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.
    Figure Legend Snippet: ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.

    Techniques Used: Translocation Assay, Two Tailed Test

    ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.
    Figure Legend Snippet: ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.

    Techniques Used:

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    Cell Signaling Technology Inc total akt2
    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, <t>p-Akt2,</t> and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
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    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, <t>p-Akt2,</t> and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
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    Cell Signaling Technology Inc ser 474
    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, <t>p-Akt2,</t> and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.
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    Cell Signaling Technology Inc phospho akt2 s473
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    Cell Signaling Technology Inc p akt2
    Composition of Akt isoforms in primary human subcutaneous preadipocytes. ( A – D ) Electropherograms of capillary isoelectric focusing (cIEF) immunoassays of ( A ) Akt1, ( B ) <t>Akt2,</t> ( C ) Akt3, and ( D ) pan-Akt or all Akt isoforms. The letter p represents phosphorylation of Akt isoforms. Peaks on electropherograms are color-coded with Akt1 (blue), Akt2 (green), and Akt3 (red). ( E ) Percentage of Akt1 (blue), Akt2 (green), and Akt3 (red) as a function of all Akt isoforms. ( F – H ) Electropherograms of cIEF immunoassays of ( F ) pAkt (Thr308), ( G ) pAkt (Thr450), and ( H ) pAkt (S473). The dashed line highlights the distribution of pAkt (Thr450) as a function of pI values.
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    Cell Signaling Technology Inc pathscan phospho akt2 ser474
    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of <t>AKT2</t> normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.
    Pathscan Phospho Akt2 Ser474, 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 akt2 levels
    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of <t>AKT2</t> normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.
    Akt2 Levels, 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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    Image Search Results


    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Journal: International Journal of Biological Sciences

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    doi: 10.7150/ijbs.12101

    Figure Lengend Snippet: Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Article Snippet: AMP-activated protein kinase α (AMPKα), phospho-AMPKα (Thr172), phospho-Akt (Ser473, p -Akt), total Akt ( t -Akt), phospho-Akt2 (Ser474, p -Akt2), and total Akt2 ( t -Akt2) were all obtained from Cell Signaling Technology (Danvers, MA), and 3-nitrotyrosine (3-NT) was purchased from Millipore (Billerica, MA).

    Techniques: Expressing, Western Blot

    Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Journal: International Journal of Biological Sciences

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    doi: 10.7150/ijbs.12101

    Figure Lengend Snippet: Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Article Snippet: AMP-activated protein kinase α (AMPKα), phospho-AMPKα (Thr172), phospho-Akt (Ser473, p -Akt), total Akt ( t -Akt), phospho-Akt2 (Ser474, p -Akt2), and total Akt2 ( t -Akt2) were all obtained from Cell Signaling Technology (Danvers, MA), and 3-nitrotyrosine (3-NT) was purchased from Millipore (Billerica, MA).

    Techniques: Expressing, Western Blot

    A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Journal: International Journal of Biological Sciences

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    doi: 10.7150/ijbs.12101

    Figure Lengend Snippet: A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Article Snippet: AMP-activated protein kinase α (AMPKα), phospho-AMPKα (Thr172), phospho-Akt (Ser473, p -Akt), total Akt ( t -Akt), phospho-Akt2 (Ser474, p -Akt2), and total Akt2 ( t -Akt2) were all obtained from Cell Signaling Technology (Danvers, MA), and 3-nitrotyrosine (3-NT) was purchased from Millipore (Billerica, MA).

    Techniques: Activity Assay, Expressing

    Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Journal: International Journal of Biological Sciences

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    doi: 10.7150/ijbs.12101

    Figure Lengend Snippet: Effects of MH on cardiac glucose metabolism. Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND; b, p< 0.05 vs. HFD.

    Article Snippet: AMP-activated protein kinase α (AMPKα), phospho-AMPKα (Thr172), phospho-Akt (Ser473, p -Akt), total Akt ( t -Akt), phospho-Akt2 (Ser474, p -Akt2), and total Akt2 ( t -Akt2) were all obtained from Cell Signaling Technology (Danvers, MA), and 3-nitrotyrosine (3-NT) was purchased from Millipore (Billerica, MA).

    Techniques: Expressing, Western Blot

    Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Journal: International Journal of Biological Sciences

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    doi: 10.7150/ijbs.12101

    Figure Lengend Snippet: Effects of MH on cardiac glucose metabolism after insulin treatment for 15 min. The data for insulin treatment (black column) are presented as the fold change relative to baseline (without insulin treatment, white column). Protein expression of p-Akt, p-Akt2, and HK II in ND-fed mice (A) and HFD-fed mice (B) was measured by western blot analysis. Data are presented as means ± SD (n=5). a, p<0.05 vs. ND+Insulin; b, p< 0.05 vs. HFD+Insulin; c, p<0.05 vs. HFD/BL153+Insulin.

    Article Snippet: AMP-activated protein kinase α (AMPKα), phospho-AMPKα (Thr172), phospho-Akt (Ser473, p -Akt), total Akt ( t -Akt), phospho-Akt2 (Ser474, p -Akt2), and total Akt2 ( t -Akt2) were all obtained from Cell Signaling Technology (Danvers, MA), and 3-nitrotyrosine (3-NT) was purchased from Millipore (Billerica, MA).

    Techniques: Expressing, Western Blot

    A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Journal: International Journal of Biological Sciences

    Article Title: Magnolia Bioactive Constituent 4-O-Methylhonokiol Prevents the Impairment of Cardiac Insulin Signaling and the Cardiac Pathogenesis in High-Fat Diet-Induced Obese Mice

    doi: 10.7150/ijbs.12101

    Figure Lengend Snippet: A proposed mechanism for the effects of MH on cardiac insulin resistance in obese mice. MH might upregulate Akt (mainly Akt2) activity to ameliorate obesity-impaired cardiac insulin signaling. On the other hand, MH might attenuate oxidative stress-induced insulin resistance by activating the Akt-Nrf2 pathway. Lastly, MH might inhibit expression of CD36, which decreases FA uptake, and preserve the FA oxidation pathway (AMPK-Sirt1-PGC-1α), subsequently reducing intracellular lipid accumulation and finally reversing insulin signaling.

    Article Snippet: AMP-activated protein kinase α (AMPKα), phospho-AMPKα (Thr172), phospho-Akt (Ser473, p -Akt), total Akt ( t -Akt), phospho-Akt2 (Ser474, p -Akt2), and total Akt2 ( t -Akt2) were all obtained from Cell Signaling Technology (Danvers, MA), and 3-nitrotyrosine (3-NT) was purchased from Millipore (Billerica, MA).

    Techniques: Activity Assay, Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: phospho-AKT2(S473) , Cell Signaling Technology , Cat# 4060; RRID: AB_2224726.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Article Snippet: phospho-AKT2(S473) , Cell Signaling Technology , Cat# 4060; RRID: AB_2224726.

    Techniques: Western Blot

    (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Article Snippet: phospho-AKT2(S473) , Cell Signaling Technology , Cat# 4060; RRID: AB_2224726.

    Techniques:

    (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Article Snippet: phospho-AKT2(S473) , Cell Signaling Technology , Cat# 4060; RRID: AB_2224726.

    Techniques:

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: phospho-AKT2(S473) , Cell Signaling Technology , Cat# 4060; RRID: AB_2224726.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: phospho-AKT2(T308) , Cell Signaling Technology , Cat# 13038; RRID: AB_2629447.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: (A) DHA and EPA specifically promote membrane-targeting proteins to the membrane. HepG2 cells were treated with DHA or EPA for 1 h before their membrane fractions were obtained and detected for AKT2, PDK1, TRAF6, SIRT3, COX4, PGK1, GAPDH, and Na-K-ATPase by western blotting.

    Article Snippet: phospho-AKT2(T308) , Cell Signaling Technology , Cat# 13038; RRID: AB_2629447.

    Techniques: Western Blot

    (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: (A) DHA and EPA activate PDK1 and AKT2. Phosphorylation levels of substrates of PDK1 and AKT2 were detected in 3T3-L1 cells treated with DHA and EPA.

    Article Snippet: phospho-AKT2(T308) , Cell Signaling Technology , Cat# 13038; RRID: AB_2629447.

    Techniques:

    (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: (A) DHA acylation induced insulin resistance. Insulin tolerance tests were performed in DHA-treated Akt2−/− 129/C57BL6 mice and Akt2−/− 129/C57BL6 mice overexpressing AKT2WT or AKT2W414L (n = 6, mean ± SEM).

    Article Snippet: phospho-AKT2(T308) , Cell Signaling Technology , Cat# 13038; RRID: AB_2629447.

    Techniques:

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Methylene-bridge tryptophan fatty acylation regulates PI3K-AKT signaling and glucose uptake

    doi: 10.1016/j.celrep.2022.110509

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: phospho-AKT2(T308) , Cell Signaling Technology , Cat# 13038; RRID: AB_2629447.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Kinase Assay, Software

    Composition of Akt isoforms in primary human subcutaneous preadipocytes. ( A – D ) Electropherograms of capillary isoelectric focusing (cIEF) immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, and ( D ) pan-Akt or all Akt isoforms. The letter p represents phosphorylation of Akt isoforms. Peaks on electropherograms are color-coded with Akt1 (blue), Akt2 (green), and Akt3 (red). ( E ) Percentage of Akt1 (blue), Akt2 (green), and Akt3 (red) as a function of all Akt isoforms. ( F – H ) Electropherograms of cIEF immunoassays of ( F ) pAkt (Thr308), ( G ) pAkt (Thr450), and ( H ) pAkt (S473). The dashed line highlights the distribution of pAkt (Thr450) as a function of pI values.

    Journal: Nutrients

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    doi: 10.3390/nu14163301

    Figure Lengend Snippet: Composition of Akt isoforms in primary human subcutaneous preadipocytes. ( A – D ) Electropherograms of capillary isoelectric focusing (cIEF) immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, and ( D ) pan-Akt or all Akt isoforms. The letter p represents phosphorylation of Akt isoforms. Peaks on electropherograms are color-coded with Akt1 (blue), Akt2 (green), and Akt3 (red). ( E ) Percentage of Akt1 (blue), Akt2 (green), and Akt3 (red) as a function of all Akt isoforms. ( F – H ) Electropherograms of cIEF immunoassays of ( F ) pAkt (Thr308), ( G ) pAkt (Thr450), and ( H ) pAkt (S473). The dashed line highlights the distribution of pAkt (Thr450) as a function of pI values.

    Article Snippet: Primary antibodies used for this study were: pan-Akt (cat. no. 8312, Santa Cruz Biotech, Dallas, TX, USA), Akt 1 (cat. no. 2938, cat. no. 2938, Cell Signaling Technology (CST), Danvers, MA, USA), Akt2 (cat. no. cs3063, CST), Akt3 (cat. no. 8018, CST), p-Akt (S473) (cat. no. 9271, CST), p-Akt1 (S473) (cat. no. 9018, CST), p-Akt2 (S474) (cat. no. cs8599, CST), p-Akt (S124) (cat. no. PA5-38251, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (S246) (cat. no. PA5-38352, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (T308) (cat. no. cs2965, CST), p-Akt (T450) (cat. no. cs9267, CST), p-Akt (Y475) (p-Akt1 (Y474) or p-Akt2 (Y475), cat. no. PA5-38351, Thermo Fisher Scientific, Waltham, MA, USA), HSP70 (cat. no. cs4872, CST), and β-actin (cat. no. MAB8929, R&D Systems, Minneapolis, MN, USA).

    Techniques:

    Insulin induces changes to the electropherograms of Akt2 and Akt3. Electropherograms of cIEF immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, ( D ) p-Akt (Thr308), ( E ) p-Akt (Thr450), and ( F ) p-Akt (Ser473) following treatment with insulin for 30 min. Arrows point to new peaks that appeared after treatment with insulin. The dashed line highlights the distribution of p-Akt (Thr450) as a function of pI values. ( G ) Capillary Western (CW) immunoassays of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before and after treatment with insulin. Pan-Akt and β-actin served as the loading controls. ( H ) Relative expression level of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before (solid black) and after (textured) treatment with insulin. Error bars indicate standard deviations across six repeated measurements using CW immunoassays per experimental condition. Asterisks indicate a statistical significance of p ≤ 0.01 versus untreated control.

    Journal: Nutrients

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    doi: 10.3390/nu14163301

    Figure Lengend Snippet: Insulin induces changes to the electropherograms of Akt2 and Akt3. Electropherograms of cIEF immunoassays of ( A ) Akt1, ( B ) Akt2, ( C ) Akt3, ( D ) p-Akt (Thr308), ( E ) p-Akt (Thr450), and ( F ) p-Akt (Ser473) following treatment with insulin for 30 min. Arrows point to new peaks that appeared after treatment with insulin. The dashed line highlights the distribution of p-Akt (Thr450) as a function of pI values. ( G ) Capillary Western (CW) immunoassays of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before and after treatment with insulin. Pan-Akt and β-actin served as the loading controls. ( H ) Relative expression level of p-Akt (Thr308), p-Akt (Thr450), and p-Akt (Ser473) before (solid black) and after (textured) treatment with insulin. Error bars indicate standard deviations across six repeated measurements using CW immunoassays per experimental condition. Asterisks indicate a statistical significance of p ≤ 0.01 versus untreated control.

    Article Snippet: Primary antibodies used for this study were: pan-Akt (cat. no. 8312, Santa Cruz Biotech, Dallas, TX, USA), Akt 1 (cat. no. 2938, cat. no. 2938, Cell Signaling Technology (CST), Danvers, MA, USA), Akt2 (cat. no. cs3063, CST), Akt3 (cat. no. 8018, CST), p-Akt (S473) (cat. no. 9271, CST), p-Akt1 (S473) (cat. no. 9018, CST), p-Akt2 (S474) (cat. no. cs8599, CST), p-Akt (S124) (cat. no. PA5-38251, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (S246) (cat. no. PA5-38352, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (T308) (cat. no. cs2965, CST), p-Akt (T450) (cat. no. cs9267, CST), p-Akt (Y475) (p-Akt1 (Y474) or p-Akt2 (Y475), cat. no. PA5-38351, Thermo Fisher Scientific, Waltham, MA, USA), HSP70 (cat. no. cs4872, CST), and β-actin (cat. no. MAB8929, R&D Systems, Minneapolis, MN, USA).

    Techniques: Western Blot, Expressing

    Cinnamaldehyde and curcumin induce changes to Akt2 posttranslational modification. ( A – E ) Distribution of Akt2 as a function of isoelectric points in ( A ) control untreated preadipocytes, or ( B – E ) preadipocytes treated with ( B ) insulin, ( C ) cinnamaldehyde, ( D ) curcumin, or ( E ) combined cinnamaldehyde and curcumin. ( F ) Distribution of p-Akt2 (S474) as a function of isoelectric points in untreated control preadipocytes (top electropherogram), or preadipocytes treated with insulin (second electropherogram), cinnamaldehyde (third electropherogram), curcumin (fourth electropherogram), and combined cinnamaldehyde and curcumin (bottom electropherogram). Arrows point to the appearance of new peaks following treatment versus untreated control.

    Journal: Nutrients

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    doi: 10.3390/nu14163301

    Figure Lengend Snippet: Cinnamaldehyde and curcumin induce changes to Akt2 posttranslational modification. ( A – E ) Distribution of Akt2 as a function of isoelectric points in ( A ) control untreated preadipocytes, or ( B – E ) preadipocytes treated with ( B ) insulin, ( C ) cinnamaldehyde, ( D ) curcumin, or ( E ) combined cinnamaldehyde and curcumin. ( F ) Distribution of p-Akt2 (S474) as a function of isoelectric points in untreated control preadipocytes (top electropherogram), or preadipocytes treated with insulin (second electropherogram), cinnamaldehyde (third electropherogram), curcumin (fourth electropherogram), and combined cinnamaldehyde and curcumin (bottom electropherogram). Arrows point to the appearance of new peaks following treatment versus untreated control.

    Article Snippet: Primary antibodies used for this study were: pan-Akt (cat. no. 8312, Santa Cruz Biotech, Dallas, TX, USA), Akt 1 (cat. no. 2938, cat. no. 2938, Cell Signaling Technology (CST), Danvers, MA, USA), Akt2 (cat. no. cs3063, CST), Akt3 (cat. no. 8018, CST), p-Akt (S473) (cat. no. 9271, CST), p-Akt1 (S473) (cat. no. 9018, CST), p-Akt2 (S474) (cat. no. cs8599, CST), p-Akt (S124) (cat. no. PA5-38251, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (S246) (cat. no. PA5-38352, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (T308) (cat. no. cs2965, CST), p-Akt (T450) (cat. no. cs9267, CST), p-Akt (Y475) (p-Akt1 (Y474) or p-Akt2 (Y475), cat. no. PA5-38351, Thermo Fisher Scientific, Waltham, MA, USA), HSP70 (cat. no. cs4872, CST), and β-actin (cat. no. MAB8929, R&D Systems, Minneapolis, MN, USA).

    Techniques: Modification

    Cinnamaldehyde and curcumin enhance insulin-stimulated activation of Akt2. ( A – D ) Distribution of Akt2 as a function of isoelectric points in preadipocytes treated with ( A ) insulin alone, ( B ) insulin and cinnamaldehyde, ( C ) insulin and curcumin, or ( D ) insulin, cinnamaldehyde and curcumin. ( E ) Distribution of p-Akt2 (S474) as a function of isoelectric points in preadipocytes treated with insulin alone (top electropherogram), insulin and cinnamaldehyde (second electropherogram), insulin and curcumin (third electropherogram), or insulin, cinnamaldehyde, and curcumin (bottom electropherogram). ( F ) Relative abundance of p-Akt2 (S474) as a function of treatment condition. Error bars are standard deviations across nine repeated measurements. Single asterisk (*) indicates p -value ≤ 0.01 versus treatment with insulin alone. Double asterisk (**) indicates p -value ≤ 0.01 versus treatment with insulin and cinnamaldehyde or insulin and curcumin.

    Journal: Nutrients

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    doi: 10.3390/nu14163301

    Figure Lengend Snippet: Cinnamaldehyde and curcumin enhance insulin-stimulated activation of Akt2. ( A – D ) Distribution of Akt2 as a function of isoelectric points in preadipocytes treated with ( A ) insulin alone, ( B ) insulin and cinnamaldehyde, ( C ) insulin and curcumin, or ( D ) insulin, cinnamaldehyde and curcumin. ( E ) Distribution of p-Akt2 (S474) as a function of isoelectric points in preadipocytes treated with insulin alone (top electropherogram), insulin and cinnamaldehyde (second electropherogram), insulin and curcumin (third electropherogram), or insulin, cinnamaldehyde, and curcumin (bottom electropherogram). ( F ) Relative abundance of p-Akt2 (S474) as a function of treatment condition. Error bars are standard deviations across nine repeated measurements. Single asterisk (*) indicates p -value ≤ 0.01 versus treatment with insulin alone. Double asterisk (**) indicates p -value ≤ 0.01 versus treatment with insulin and cinnamaldehyde or insulin and curcumin.

    Article Snippet: Primary antibodies used for this study were: pan-Akt (cat. no. 8312, Santa Cruz Biotech, Dallas, TX, USA), Akt 1 (cat. no. 2938, cat. no. 2938, Cell Signaling Technology (CST), Danvers, MA, USA), Akt2 (cat. no. cs3063, CST), Akt3 (cat. no. 8018, CST), p-Akt (S473) (cat. no. 9271, CST), p-Akt1 (S473) (cat. no. 9018, CST), p-Akt2 (S474) (cat. no. cs8599, CST), p-Akt (S124) (cat. no. PA5-38251, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (S246) (cat. no. PA5-38352, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (T308) (cat. no. cs2965, CST), p-Akt (T450) (cat. no. cs9267, CST), p-Akt (Y475) (p-Akt1 (Y474) or p-Akt2 (Y475), cat. no. PA5-38351, Thermo Fisher Scientific, Waltham, MA, USA), HSP70 (cat. no. cs4872, CST), and β-actin (cat. no. MAB8929, R&D Systems, Minneapolis, MN, USA).

    Techniques: Activation Assay

    Cinnamaldehyde and curcumin increase pan-Akt phosphorylation. ( A – C ) Distribution of p-Akt (T450) as a function of isoelectric points in ( A ) untreated control preadipocytes or ( B , C ) preadipocytes treated with ( B ) cinnamaldehyde or ( C ) curcumin. The dashed line highlights the distribution trend. ( D – F ) Distribution of ( D ) p-Akt (S124), ( E ) p-Akt (S246), and ( F ) p-Akt (Y475) in untreated control preadipocytes (top electropherogram), or preadipocytes treated with cinnamaldehyde (middle electropherogram) or curcumin (bottom electropherogram).

    Journal: Nutrients

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    doi: 10.3390/nu14163301

    Figure Lengend Snippet: Cinnamaldehyde and curcumin increase pan-Akt phosphorylation. ( A – C ) Distribution of p-Akt (T450) as a function of isoelectric points in ( A ) untreated control preadipocytes or ( B , C ) preadipocytes treated with ( B ) cinnamaldehyde or ( C ) curcumin. The dashed line highlights the distribution trend. ( D – F ) Distribution of ( D ) p-Akt (S124), ( E ) p-Akt (S246), and ( F ) p-Akt (Y475) in untreated control preadipocytes (top electropherogram), or preadipocytes treated with cinnamaldehyde (middle electropherogram) or curcumin (bottom electropherogram).

    Article Snippet: Primary antibodies used for this study were: pan-Akt (cat. no. 8312, Santa Cruz Biotech, Dallas, TX, USA), Akt 1 (cat. no. 2938, cat. no. 2938, Cell Signaling Technology (CST), Danvers, MA, USA), Akt2 (cat. no. cs3063, CST), Akt3 (cat. no. 8018, CST), p-Akt (S473) (cat. no. 9271, CST), p-Akt1 (S473) (cat. no. 9018, CST), p-Akt2 (S474) (cat. no. cs8599, CST), p-Akt (S124) (cat. no. PA5-38251, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (S246) (cat. no. PA5-38352, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (T308) (cat. no. cs2965, CST), p-Akt (T450) (cat. no. cs9267, CST), p-Akt (Y475) (p-Akt1 (Y474) or p-Akt2 (Y475), cat. no. PA5-38351, Thermo Fisher Scientific, Waltham, MA, USA), HSP70 (cat. no. cs4872, CST), and β-actin (cat. no. MAB8929, R&D Systems, Minneapolis, MN, USA).

    Techniques:

    Dual inhibition of PP2A and PTP1B activates Akt2. ( A – C ) Distribution of Akt2 as a function of isoelectric points in control untreated preadipocytes (top electropherograms) or in preadipocytes treated with ( A ) okadaic acid, ( B ) PTP1Bi, or ( C ) combined okadaic acid and PTP1B (bottom electropherograms). Arrows point to new peaks or peaks that experience elevated abundance in treated preadipocytes versus untreated control preadipocytes. ( D ) Distribution of p-Akt2 (S474) as a function of isoelectric point in control untreated preadipocytes (top electropherogram), or preadipocytes treated with okadaic acid (second electropherogram), PTP1Bi (third electropherogram), or combined okadaic acid and PTP1Bi (bottom electropherogram).

    Journal: Nutrients

    Article Title: Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation

    doi: 10.3390/nu14163301

    Figure Lengend Snippet: Dual inhibition of PP2A and PTP1B activates Akt2. ( A – C ) Distribution of Akt2 as a function of isoelectric points in control untreated preadipocytes (top electropherograms) or in preadipocytes treated with ( A ) okadaic acid, ( B ) PTP1Bi, or ( C ) combined okadaic acid and PTP1B (bottom electropherograms). Arrows point to new peaks or peaks that experience elevated abundance in treated preadipocytes versus untreated control preadipocytes. ( D ) Distribution of p-Akt2 (S474) as a function of isoelectric point in control untreated preadipocytes (top electropherogram), or preadipocytes treated with okadaic acid (second electropherogram), PTP1Bi (third electropherogram), or combined okadaic acid and PTP1Bi (bottom electropherogram).

    Article Snippet: Primary antibodies used for this study were: pan-Akt (cat. no. 8312, Santa Cruz Biotech, Dallas, TX, USA), Akt 1 (cat. no. 2938, cat. no. 2938, Cell Signaling Technology (CST), Danvers, MA, USA), Akt2 (cat. no. cs3063, CST), Akt3 (cat. no. 8018, CST), p-Akt (S473) (cat. no. 9271, CST), p-Akt1 (S473) (cat. no. 9018, CST), p-Akt2 (S474) (cat. no. cs8599, CST), p-Akt (S124) (cat. no. PA5-38251, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (S246) (cat. no. PA5-38352, Thermo Fisher Scientific, Waltham, MA, USA), p-Akt (T308) (cat. no. cs2965, CST), p-Akt (T450) (cat. no. cs9267, CST), p-Akt (Y475) (p-Akt1 (Y474) or p-Akt2 (Y475), cat. no. PA5-38351, Thermo Fisher Scientific, Waltham, MA, USA), HSP70 (cat. no. cs4872, CST), and β-actin (cat. no. MAB8929, R&D Systems, Minneapolis, MN, USA).

    Techniques: Inhibition

    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.

    Journal: Science Advances

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    doi: 10.1126/sciadv.abn7298

    Figure Lengend Snippet: ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.

    Article Snippet: PathScan Phospho-Akt2 (Ser474) and Total Akt2 Sandwich ELISA kits were used to quantify AKT2 levels (Cell Signaling Technology, #7048 and #7046, respectively) as per the manufacturer’s instructions, by colorimetric reading at 450 nm on a Thermo Fisher Multiskan Go plate reader.

    Techniques: Functional Assay

    ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.

    Journal: Science Advances

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    doi: 10.1126/sciadv.abn7298

    Figure Lengend Snippet: ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.

    Article Snippet: PathScan Phospho-Akt2 (Ser474) and Total Akt2 Sandwich ELISA kits were used to quantify AKT2 levels (Cell Signaling Technology, #7048 and #7046, respectively) as per the manufacturer’s instructions, by colorimetric reading at 450 nm on a Thermo Fisher Multiskan Go plate reader.

    Techniques: Translocation Assay, Two Tailed Test

    ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.

    Journal: Science Advances

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    doi: 10.1126/sciadv.abn7298

    Figure Lengend Snippet: ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.

    Article Snippet: PathScan Phospho-Akt2 (Ser474) and Total Akt2 Sandwich ELISA kits were used to quantify AKT2 levels (Cell Signaling Technology, #7048 and #7046, respectively) as per the manufacturer’s instructions, by colorimetric reading at 450 nm on a Thermo Fisher Multiskan Go plate reader.

    Techniques:

    ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.

    Journal: Science Advances

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    doi: 10.1126/sciadv.abn7298

    Figure Lengend Snippet: ( A ) Schematic indicating the experimental setup to potentiate insulin response in functional assays. hPSCs are differentiated into adipocytes using a published protocol, after which an additional step was added and optimized to sensitize adipocytes. ( B and C ) Phosphorylation of AKT2 normalized to total AKT2 (B) and glucose uptake (C) after 100 nM insulin stimulation measured for all the medium compositions from our DoE model and the medium from a previously published protocol. RLU, relative light unit. ( D and E ) The parameter coefficient of each factor in our DoE model indicating its contribution to the AKT2 phosphorylation (D) and glucose uptake (E). ( F ) Time course of sensitization with the DoE-optimized media measuring glucose uptake at baseline and after insulin stimulation. Results are normalized to total protein for each sample. All bar graphs depict the mean with error bars representing SD, n = 2 biological replicates. OD 450 , optical density at 450 nm.

    Article Snippet: PathScan Phospho-Akt2 (Ser474) and Total Akt2 Sandwich ELISA kits were used to quantify AKT2 levels (Cell Signaling Technology, #7048 and #7046, respectively) as per the manufacturer’s instructions, by colorimetric reading at 450 nm on a Thermo Fisher Multiskan Go plate reader.

    Techniques: Functional Assay

    ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.

    Journal: Science Advances

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    doi: 10.1126/sciadv.abn7298

    Figure Lengend Snippet: ( A ) Schematic indicating the experimental setup to measure insulin dose-response and induction of insulin resistance. ( B ) Insulin dose-response curve showing fold change in AKT2 phosphorylation compared to the unstimulated state. ( C ) Representative images of GLUT4 translocation to the cell membrane upon insulin stimulation. ( D ) TIRF measurement of GLUT4 signal intensity at the adipocyte cell membrane (** P < 0.01, unpaired two-tailed t test). ( E ) Insulin dose-response curve showing AKT2 phosphorylation fold change in three insulin preexposure conditions. Results are normalized to total AKT2 and plotted as fold change to unstimulated cells in that condition. ( F ) Insulin dose-response curve showing glucose uptake for sensitized and hyperinsulinemia-exposed adipocytes. Results are normalized to total protein content and plotted as fold change to unstimulated cells. ( G ) Glycerol release into the medium for published, sensitized, and hyperinsulinemia-exposed adipocytes showing basal or insulin-suppressed lipolysis, normalized to total protein. Bar graphs depict the mean with error bars representing SD, dose-response curves depict a nonlinear fit curve with error bars representing SD, and the scatterplot depicts individual cell values with mean and 95% confidence interval (CI) overlaid; n = 3 biological replicates unless otherwise indicated.

    Article Snippet: PathScan Phospho-Akt2 (Ser474) and Total Akt2 Sandwich ELISA kits were used to quantify AKT2 levels (Cell Signaling Technology, #7048 and #7046, respectively) as per the manufacturer’s instructions, by colorimetric reading at 450 nm on a Thermo Fisher Multiskan Go plate reader.

    Techniques: Translocation Assay, Two Tailed Test

    ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.

    Journal: Science Advances

    Article Title: Development of a physiological insulin resistance model in human stem cell–derived adipocytes

    doi: 10.1126/sciadv.abn7298

    Figure Lengend Snippet: ( A ) Phosphorylated AKT2 measurements as a fraction of total AKT2 normalized to total protein per well for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. ( B ) Glucose uptake measurements normalized to total protein per well and displayed as fold change in glucose uptake versus the unstimulated condition for brown hPSC adipocytes, mouse 3T3-L1 differentiated adipocytes, and primary human SVF adipocytes. All bar graphs depict the mean with error bars representing SD, n = 4 biological replicates.

    Article Snippet: PathScan Phospho-Akt2 (Ser474) and Total Akt2 Sandwich ELISA kits were used to quantify AKT2 levels (Cell Signaling Technology, #7048 and #7046, respectively) as per the manufacturer’s instructions, by colorimetric reading at 450 nm on a Thermo Fisher Multiskan Go plate reader.

    Techniques: