rabbit anti c terminal herg antibody  (Alomone Labs)


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    Alomone Labs rabbit anti c terminal herg antibody
    (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of <t>HERG</t> 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with <t>anti-C</t> terminal HERG antibody and were repeated twice (n = 2).
    Rabbit Anti C Terminal Herg Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti c terminal herg antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti c terminal herg antibody - by Bioz Stars, 2024-07
    93/100 stars

    Images

    1) Product Images from "Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel"

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0032654

    (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).
    Figure Legend Snippet: (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Techniques Used: Western Blot, Expressing, Mutagenesis

    (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).
    Figure Legend Snippet: (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Techniques Used: Western Blot, Transfection, Expressing, Mutagenesis, Bicinchoninic Acid Protein Assay

    (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).
    Figure Legend Snippet: (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Techniques Used: Western Blot, Expressing

    rat herg  (Alomone Labs)


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    Alomone Labs rat herg
    LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses <t>of</t> <t>ATF4,</t> ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, <t>HERG,</t> SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test
    Rat Herg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat herg/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rat herg - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "LOC102549726/miR-760-3p network is involved in the progression of ISO-induced pathological cardiomyocyte hypertrophy via endoplasmic reticulum stress"

    Article Title: LOC102549726/miR-760-3p network is involved in the progression of ISO-induced pathological cardiomyocyte hypertrophy via endoplasmic reticulum stress

    Journal: Journal of Molecular Histology

    doi: 10.1007/s10735-023-10166-1

    LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, HERG, SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test
    Figure Legend Snippet: LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, HERG, SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test

    Techniques Used: Synthesized, Real-time Polymerase Chain Reaction, Staining, Transfection, Negative Control, Western Blot, Over Expression, Expressing

    Roles of miR-760-3p in mediating the anti-hypertrophic effect in ISO-induced cardiomyocyte hypertrophy via ER stress. ( a ) The miRNA relative expression level of synthesized miR-760-3p mimic and miR-760-3p inhibitor were verified using qPCR. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b,c ) Representative H&E staining images of cardiomyocytes and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. Transfection with miR-760-3p inhibitor resulted in larger cell size than NC transfection, whereas miR-760-3p mimic significantly decreased cell surface area in NRCMs. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( d-f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1, HERG, SERCA2 and BNP in NRCMs transfected with miR-760-3p mimic and inhibitor upon ISO treatment in the indicated groups. MiR-760-3p inhibitor enhanced the expression of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1 and BNP, while simultaneously reducing p-eIF2α, p-eIF2α/eIF2α ratio, HERG, and SERCA2. Conversely, transfection with miR-760-3p mimic showed the opposite effect. Bars show the mean values ± SD. n = 3, *p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test
    Figure Legend Snippet: Roles of miR-760-3p in mediating the anti-hypertrophic effect in ISO-induced cardiomyocyte hypertrophy via ER stress. ( a ) The miRNA relative expression level of synthesized miR-760-3p mimic and miR-760-3p inhibitor were verified using qPCR. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b,c ) Representative H&E staining images of cardiomyocytes and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. Transfection with miR-760-3p inhibitor resulted in larger cell size than NC transfection, whereas miR-760-3p mimic significantly decreased cell surface area in NRCMs. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( d-f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1, HERG, SERCA2 and BNP in NRCMs transfected with miR-760-3p mimic and inhibitor upon ISO treatment in the indicated groups. MiR-760-3p inhibitor enhanced the expression of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1 and BNP, while simultaneously reducing p-eIF2α, p-eIF2α/eIF2α ratio, HERG, and SERCA2. Conversely, transfection with miR-760-3p mimic showed the opposite effect. Bars show the mean values ± SD. n = 3, *p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test

    Techniques Used: Expressing, Synthesized, Staining, Transfection, Western Blot

    rabbit polyclonal anti kcnh2 herg antibody  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti kcnh2 herg antibody
    Rabbit Polyclonal Anti Kcnh2 Herg Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti kcnh2 herg antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti kcnh2 herg antibody - by Bioz Stars, 2024-07
    86/100 stars

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    rabbit polyclonal anti kcnh2 herg antibody  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti kcnh2 herg antibody
    Rabbit Polyclonal Anti Kcnh2 Herg Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti kcnh2 herg antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti kcnh2 herg antibody - by Bioz Stars, 2024-07
    86/100 stars

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    anti kv11 1 herg  (Alomone Labs)


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    Alomone Labs anti kv11 1 herg
    Anti Kv11 1 Herg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti kv11 1 herg/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti kv11 1 herg - by Bioz Stars, 2024-07
    86/100 stars

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    herg primary antibody  (Alomone Labs)


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    Alomone Labs herg primary antibody
    Herg Primary Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/herg primary antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    herg primary antibody - by Bioz Stars, 2024-07
    86/100 stars

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    herg primary antibody  (Alomone Labs)


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    Alomone Labs herg primary antibody
    Herg Primary Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/herg primary antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    herg primary antibody - by Bioz Stars, 2024-07
    86/100 stars

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    anti herg  (Alomone Labs)


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    Alomone Labs anti herg
    Anti Herg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti herg/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti herg - by Bioz Stars, 2024-07
    86/100 stars

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    rabbit anti c terminal herg antibody  (Alomone Labs)


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    Alomone Labs rabbit anti c terminal herg antibody
    (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of <t>HERG</t> 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with <t>anti-C</t> terminal HERG antibody and were repeated twice (n = 2).
    Rabbit Anti C Terminal Herg Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti c terminal herg antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti c terminal herg antibody - by Bioz Stars, 2024-07
    93/100 stars

    Images

    1) Product Images from "Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel"

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0032654

    (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).
    Figure Legend Snippet: (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Techniques Used: Western Blot, Expressing, Mutagenesis

    (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).
    Figure Legend Snippet: (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Techniques Used: Western Blot, Transfection, Expressing, Mutagenesis, Bicinchoninic Acid Protein Assay

    (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).
    Figure Legend Snippet: (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Techniques Used: Western Blot, Expressing

    anti kcnh2  (Alomone Labs)


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    Alomone Labs anti kcnh2
    Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG <t>(KCNH2)</t> protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.
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    Images

    1) Product Images from "Stress-induced premature senescence is associated with a prolonged QT interval and recapitulates features of cardiac aging"

    Article Title: Stress-induced premature senescence is associated with a prolonged QT interval and recapitulates features of cardiac aging

    Journal: Theranostics

    doi: 10.7150/thno.70884

    Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG (KCNH2) protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.
    Figure Legend Snippet: Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG (KCNH2) protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.

    Techniques Used: Expressing

    apc-109  (Alomone Labs)


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    Alomone Labs apc-109
    Apc 109, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit anti c terminal herg antibody
    (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of <t>HERG</t> 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with <t>anti-C</t> terminal HERG antibody and were repeated twice (n = 2).
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    LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses <t>of</t> <t>ATF4,</t> ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, <t>HERG,</t> SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test
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    LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses <t>of</t> <t>ATF4,</t> ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, <t>HERG,</t> SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test
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    LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses <t>of</t> <t>ATF4,</t> ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, <t>HERG,</t> SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test
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    Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG <t>(KCNH2)</t> protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.
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    Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG <t>(KCNH2)</t> protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.
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    Image Search Results


    (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Journal: PLoS ONE

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    doi: 10.1371/journal.pone.0032654

    Figure Lengend Snippet: (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Article Snippet: Proteins were transferred by semi-dry blotting to a nitrocellulose membrane and incubated overnight at 4°C with a rabbit anti-C terminal HERG antibody (Alomone Labs) and detected by ECL.

    Techniques: Western Blot, Expressing, Mutagenesis

    (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Journal: PLoS ONE

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    doi: 10.1371/journal.pone.0032654

    Figure Lengend Snippet: (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Article Snippet: Proteins were transferred by semi-dry blotting to a nitrocellulose membrane and incubated overnight at 4°C with a rabbit anti-C terminal HERG antibody (Alomone Labs) and detected by ECL.

    Techniques: Western Blot, Transfection, Expressing, Mutagenesis, Bicinchoninic Acid Protein Assay

    (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Journal: PLoS ONE

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    doi: 10.1371/journal.pone.0032654

    Figure Lengend Snippet: (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Article Snippet: Proteins were transferred by semi-dry blotting to a nitrocellulose membrane and incubated overnight at 4°C with a rabbit anti-C terminal HERG antibody (Alomone Labs) and detected by ECL.

    Techniques: Western Blot, Expressing

    LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, HERG, SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test

    Journal: Journal of Molecular Histology

    Article Title: LOC102549726/miR-760-3p network is involved in the progression of ISO-induced pathological cardiomyocyte hypertrophy via endoplasmic reticulum stress

    doi: 10.1007/s10735-023-10166-1

    Figure Lengend Snippet: LOC102549726 promotes CH via ER stress in ISO-treated NRCMs. ( a ) The interfering effect of the three synthesized siRNA-LOC102549726 was evaluated and compared with the NC group by real-time PCR, and Si-435 was selected for subsequent experiments. n = 3 in each group. Values are expressed as mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b ) Knockdown of LOC102549726 by siRNA reversed the ISO-induced increase in mRNA levels of ANP, ATF 6 induced by ISO in NRCMs. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. control group; # p < 0.05, vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( c,d ) Representative H&E staining images and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. The hypertrophic response was reversed after transfection with si-LOC102549726 as compared to ISO-treated and negative control (NC) transfected groups. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( e,f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, and p-eIF2α in the indicated groups. si-LOC102549726 treatment significantly reduced ISO-induced overexpression of ATF6, eIF2α and p-eIF2α in NRCMs. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( g,h ) Representative immunoblots and quantitative analyses of p-IRE, XBP-1, HERG, SERCA2 and BNP in the indicated groups. si-LOC102549726 treatment enhanced the expression levels of HERG and SERCA2 than those in the control group, whereas the levels of p-IRE, XBP-1 and BNP reduced. Bars show the mean values ± SD. n = 3, * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test

    Article Snippet: The membranes were blocked with 5% skimmed milk in Tris-buffered saline Tween (TBST) for 1 h at room temperature and then incubated with the following primary antibodies; Rabbit anti rat ATF6 (1:1000; cat. no. ab37149; Abcam), Rabbit Anti rat p-IRE1 (1:1000; cat. no.14C10; CST), Rabbit Anti rat p-eIF2α (1:1000; cat. no. #9721; CST), Rabbit Anti rat XBP-1 (1:1000; cat. no. abs115725; absin), Rabbit Anti rat ATF4 (1:1000; cat. no. abs135528; absin), Rabbit Anti rat HERG (1:1000; cat. no. APC-062; alomone labs), and Rabbit Anti rat SERCA2 (1:1000; cat. no. 4388; CST) overnight at 4 °C.

    Techniques: Synthesized, Real-time Polymerase Chain Reaction, Staining, Transfection, Negative Control, Western Blot, Over Expression, Expressing

    Roles of miR-760-3p in mediating the anti-hypertrophic effect in ISO-induced cardiomyocyte hypertrophy via ER stress. ( a ) The miRNA relative expression level of synthesized miR-760-3p mimic and miR-760-3p inhibitor were verified using qPCR. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b,c ) Representative H&E staining images of cardiomyocytes and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. Transfection with miR-760-3p inhibitor resulted in larger cell size than NC transfection, whereas miR-760-3p mimic significantly decreased cell surface area in NRCMs. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( d-f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1, HERG, SERCA2 and BNP in NRCMs transfected with miR-760-3p mimic and inhibitor upon ISO treatment in the indicated groups. MiR-760-3p inhibitor enhanced the expression of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1 and BNP, while simultaneously reducing p-eIF2α, p-eIF2α/eIF2α ratio, HERG, and SERCA2. Conversely, transfection with miR-760-3p mimic showed the opposite effect. Bars show the mean values ± SD. n = 3, *p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test

    Journal: Journal of Molecular Histology

    Article Title: LOC102549726/miR-760-3p network is involved in the progression of ISO-induced pathological cardiomyocyte hypertrophy via endoplasmic reticulum stress

    doi: 10.1007/s10735-023-10166-1

    Figure Lengend Snippet: Roles of miR-760-3p in mediating the anti-hypertrophic effect in ISO-induced cardiomyocyte hypertrophy via ER stress. ( a ) The miRNA relative expression level of synthesized miR-760-3p mimic and miR-760-3p inhibitor were verified using qPCR. n = 3 in each group. Values are mean ± SD. * p < 0.05, vs. NC group, using an ANOVA test. ( b,c ) Representative H&E staining images of cardiomyocytes and quantitative analyses of myocardial hypertrophy based on cell surface area in the indicated groups. Transfection with miR-760-3p inhibitor resulted in larger cell size than NC transfection, whereas miR-760-3p mimic significantly decreased cell surface area in NRCMs. The datas are represented as mean ± SD, n = 3 for each group. Scale bar = 100 μm. * p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test. ( d-f ) Representative immunoblots and quantitative analyses of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1, HERG, SERCA2 and BNP in NRCMs transfected with miR-760-3p mimic and inhibitor upon ISO treatment in the indicated groups. MiR-760-3p inhibitor enhanced the expression of ATF4, ATF6, eIF2α, p-eIF2α, p-IRE, XBP-1 and BNP, while simultaneously reducing p-eIF2α, p-eIF2α/eIF2α ratio, HERG, and SERCA2. Conversely, transfection with miR-760-3p mimic showed the opposite effect. Bars show the mean values ± SD. n = 3, *p < 0.05 vs. control group; # p < 0.05 vs. ISO + NC group, respectively using Student’s t-test and one way ANOVA test

    Article Snippet: The membranes were blocked with 5% skimmed milk in Tris-buffered saline Tween (TBST) for 1 h at room temperature and then incubated with the following primary antibodies; Rabbit anti rat ATF6 (1:1000; cat. no. ab37149; Abcam), Rabbit Anti rat p-IRE1 (1:1000; cat. no.14C10; CST), Rabbit Anti rat p-eIF2α (1:1000; cat. no. #9721; CST), Rabbit Anti rat XBP-1 (1:1000; cat. no. abs115725; absin), Rabbit Anti rat ATF4 (1:1000; cat. no. abs135528; absin), Rabbit Anti rat HERG (1:1000; cat. no. APC-062; alomone labs), and Rabbit Anti rat SERCA2 (1:1000; cat. no. 4388; CST) overnight at 4 °C.

    Techniques: Expressing, Synthesized, Staining, Transfection, Western Blot

    Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG (KCNH2) protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.

    Journal: Theranostics

    Article Title: Stress-induced premature senescence is associated with a prolonged QT interval and recapitulates features of cardiac aging

    doi: 10.7150/thno.70884

    Figure Lengend Snippet: Alterations of I Kr , I NaL and Ca 2+ handling in SenCMs. (A) E-4031 (3µM)-sensitive currents (I Kr ) activated according to the protocol shown on top and relative I/V relationships in iCMs (N = 21) and SenCMs (N = 26) 5-7 days after treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (B) HERG (KCNH2) protein expression levels in iCMs and SenCMs. Quantitative data of four independent experiments ± SEM (densitometric values for the protein of interest normalized for histone H3). ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (C) TTX (2 µM)-sensitive current (I TTX ) activated during slow voltage ramps (28 mV/sec) from a holding potential of -100 mV. Mean ± SEM I/V relationships for iCMs (N = 12) and SenCMs (N = 13) are shown. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. Statistics of I TTX at 0 mV, representative of I NaL , are reported on the right. (D) pThr286 CAMKII:CAMKII total protein expression levels in SenCMs versus iCMs. Quantitative data are from four independent experiments ± SEM (densitometric values for the proteins of interest normalized for GAPDH). * P < 0.05, ** P < 0.01 vs iCMs. (E) Membrane currents and Ca 2+ transients (CaT) were recorded simultaneously according to the voltage clamp protocol shown on top in Fluo4-loaded iCMs. Examples (left panel) and statistics (right panel) of CaL influx, CaT amplitude and caffeine-induced CaT amplitude (estimating CaSR) in iCMs (N = 22) and SenCMs (N= 14) 5-7 days after Dox treatment. Data pooled from four independent experiments are presented as ± SEM. * P < 0.05, ** P < 0.01 vs iCMs. (F) SERCA2 and monomeric (m) PLN protein expression levels in iCMs and SenCMs. Quantitative data of five independent experiments ± SEM (densitometric values for the protein of interest normalized for actin). (G) Statistics of Ca 2+ spark rate and characteristics in SenCMs (N=410) vs iCMs (N=266). Line scan (xt) images are shown on the left (time bar: 100 ms). * P < 0.05, ** P < 0.01 vs iCMs.

    Article Snippet: The membranes were blocked for 1 h with Intercept (TBS) Blocking Buffer (Licor) or milk and incubated with the primary Abs at 4 °C overnight (anti-p16, 1:1'000, Proteintech 10883; anti-p21, 1:1'000 eBiosciences 14-6715-81; anti-KCNH2, 1:200, Alomone APC-109; anti-P AMPK, 1:1'000, Cell Signaling 2537; anti-CAMKII, 1:2'000, Abcam 92332; anti-P CAMKII, 1:2'000, Abcam 171095; anti-SERCA2, 1:1000, N-19 Santa Cruz Biotechnology; anti-PLN 1:1000, 2D12, Abcam; anti-actin 1:5000, Merck).

    Techniques: Expressing