anti eno1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eno1
    Anti Eno1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    enolase 1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc enolase 1
    Enolase 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    enolase 1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc enolase 1
    Enolase 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    enos 1 300 9572  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc enos 1 300 9572
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    anti eno1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eno1
    ( A ) PKM2 and <t>ENO1</t> exist in the PGAM1-associated protein complex. Top: The whole-cell lysates (WCL) from A549 cells were prepared for immunoprecipitation (IP) and analyzed by western blot (WB). Bottom: Quantification of IP. Relative levels of PKM2, ENO1, PKM1, or PCK2 in IP were respectively normalized to that in WCL for each group. ( B – D ) PGAM1 directly interacts with PKM2 but not ENO1. Recombinant His-PGAM1 was immobilized on Ni-beads and then mixed with recombinant GST-PKM2 ( B ) or GST-ENO1 ( C ) prior to PD assay. The protein complex on the Ni-beads was eluted and analyzed by WB. ( D ) Quantification of PD in ( B , C ). Relative levels of GST-PKM2 or GST-ENO1 were normalized to that of His (PGAM1) for each group. ( E ) Confirmation of PGAM1 and PKM2 interaction by BLI assay. Recombinant His-PGAM1 protein was loaded onto Ni-sensor. The sensorgrams for the interaction of PGAM1 with GST-PKM2 as well as the associated KD values are shown as indicated. ( F ) PGAM1–PKM2 interaction and PGAM1 phosphorylation are more evident in tumor cells. Left: PGAM1-associated proteins were immunoprecipitated from the WCL of different tumor cells and analyzed by WB. Right: Quantification of IP. Relative levels of PKM2 or 3-pHis were normalized to that of PGAM1 for each group. ( G ) PKM2 K270M mutation reduces PGAM1 H11 phosphorylation. Left: PKM2 WT or K270M mutants were restored in endogenous PKM2-depleted A549 cells, PGAM1-associated proteins were immunoprecipitated and analyzed by WB. Right: Quantification of IP. Relative levels of PKM2 or 3-pHis were normalized to that of PGAM1 for each group. ( H ) PKM2 directly phosphorylates PGAM1 at H11. Top: In vitro kinase assays were carried out with recombinant GST-PKM2, His-PGAM1 as well as His-PGAM1 H11N mutant in the presence of PEP under normal (pH = 8.8, unboiled), heating (pH = 8.8, boiled) or acid condition (pH = 1, unboiled). PGAM1 H11 phosphorylation was detected by WB. Bottom: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. ( I , J ) PKM2 upregulates PGAM1 activity. The PGAM1 activity ( I ) and pyruvate generation ( J ) from in vitro kinase assay in the normal condition as in ( H ) were detected by commercially available kits. ( K ) ITC binding measurements of 2,3-BPG with PGAM1. The recombinant His-PGAM1 proteins purified bacterially were injected into calorimetric cells before 2,3-BPG injection. Theoretical titration curves as well as associated KD values were displayed. ( L ) DARTS binding assay of 2,3-BPG with PGAM1. A549 cells were transfected with Flag-PGAM1 WT, Y26F or Y92F for 48 h. Top: The whole lysates were respectively incubated with the indicated doses of 2,3-BPG respectively for 30 min at room temperature followed by pronase digestion and WB analysis. Bottom: Quantification of WB. Relative levels of Flag were normalized to tubulin without pronase for each group. ( M ) PKM2 upregulates H11 phosphorylation of wild-type PGAM1 as well as Y26F and Y92F mutations. Top: A549 cells were transfected with Flag-PGAM1 WT, Y26F or Y92F for 48 h. Flag (PGAM1)-associated proteins were immunoprecipitated and analyzed by WB. Bottom: Quantification of IP. Relative levels of 3-pHis or PKM2 were normalized to that of Flag (PGAM1) for each group. Data information: for WB in ( A – C , F – H , L , M ), BLI in ( E ) and ITC in ( K ), one representative experiment out of three is shown. For IP in ( A , F , G , M ), IgG served as a negative control. Data were represented as mean ± SD ( n = 3) with significance determined by one-way ( F – J , L , M ) or two-way ( A , D ) analysis of variance (ANOVA) test; **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05. ns nonsignificant. .
    Anti Eno1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eno1/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    anti eno1 - by Bioz Stars, 2024-07
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    1) Product Images from "PKM2 functions as a histidine kinase to phosphorylate PGAM1 and increase glycolysis shunts in cancer"

    Article Title: PKM2 functions as a histidine kinase to phosphorylate PGAM1 and increase glycolysis shunts in cancer

    Journal: The EMBO Journal

    doi: 10.1038/s44318-024-00110-8

    ( A ) PKM2 and ENO1 exist in the PGAM1-associated protein complex. Top: The whole-cell lysates (WCL) from A549 cells were prepared for immunoprecipitation (IP) and analyzed by western blot (WB). Bottom: Quantification of IP. Relative levels of PKM2, ENO1, PKM1, or PCK2 in IP were respectively normalized to that in WCL for each group. ( B – D ) PGAM1 directly interacts with PKM2 but not ENO1. Recombinant His-PGAM1 was immobilized on Ni-beads and then mixed with recombinant GST-PKM2 ( B ) or GST-ENO1 ( C ) prior to PD assay. The protein complex on the Ni-beads was eluted and analyzed by WB. ( D ) Quantification of PD in ( B , C ). Relative levels of GST-PKM2 or GST-ENO1 were normalized to that of His (PGAM1) for each group. ( E ) Confirmation of PGAM1 and PKM2 interaction by BLI assay. Recombinant His-PGAM1 protein was loaded onto Ni-sensor. The sensorgrams for the interaction of PGAM1 with GST-PKM2 as well as the associated KD values are shown as indicated. ( F ) PGAM1–PKM2 interaction and PGAM1 phosphorylation are more evident in tumor cells. Left: PGAM1-associated proteins were immunoprecipitated from the WCL of different tumor cells and analyzed by WB. Right: Quantification of IP. Relative levels of PKM2 or 3-pHis were normalized to that of PGAM1 for each group. ( G ) PKM2 K270M mutation reduces PGAM1 H11 phosphorylation. Left: PKM2 WT or K270M mutants were restored in endogenous PKM2-depleted A549 cells, PGAM1-associated proteins were immunoprecipitated and analyzed by WB. Right: Quantification of IP. Relative levels of PKM2 or 3-pHis were normalized to that of PGAM1 for each group. ( H ) PKM2 directly phosphorylates PGAM1 at H11. Top: In vitro kinase assays were carried out with recombinant GST-PKM2, His-PGAM1 as well as His-PGAM1 H11N mutant in the presence of PEP under normal (pH = 8.8, unboiled), heating (pH = 8.8, boiled) or acid condition (pH = 1, unboiled). PGAM1 H11 phosphorylation was detected by WB. Bottom: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. ( I , J ) PKM2 upregulates PGAM1 activity. The PGAM1 activity ( I ) and pyruvate generation ( J ) from in vitro kinase assay in the normal condition as in ( H ) were detected by commercially available kits. ( K ) ITC binding measurements of 2,3-BPG with PGAM1. The recombinant His-PGAM1 proteins purified bacterially were injected into calorimetric cells before 2,3-BPG injection. Theoretical titration curves as well as associated KD values were displayed. ( L ) DARTS binding assay of 2,3-BPG with PGAM1. A549 cells were transfected with Flag-PGAM1 WT, Y26F or Y92F for 48 h. Top: The whole lysates were respectively incubated with the indicated doses of 2,3-BPG respectively for 30 min at room temperature followed by pronase digestion and WB analysis. Bottom: Quantification of WB. Relative levels of Flag were normalized to tubulin without pronase for each group. ( M ) PKM2 upregulates H11 phosphorylation of wild-type PGAM1 as well as Y26F and Y92F mutations. Top: A549 cells were transfected with Flag-PGAM1 WT, Y26F or Y92F for 48 h. Flag (PGAM1)-associated proteins were immunoprecipitated and analyzed by WB. Bottom: Quantification of IP. Relative levels of 3-pHis or PKM2 were normalized to that of Flag (PGAM1) for each group. Data information: for WB in ( A – C , F – H , L , M ), BLI in ( E ) and ITC in ( K ), one representative experiment out of three is shown. For IP in ( A , F , G , M ), IgG served as a negative control. Data were represented as mean ± SD ( n = 3) with significance determined by one-way ( F – J , L , M ) or two-way ( A , D ) analysis of variance (ANOVA) test; **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05. ns nonsignificant. .
    Figure Legend Snippet: ( A ) PKM2 and ENO1 exist in the PGAM1-associated protein complex. Top: The whole-cell lysates (WCL) from A549 cells were prepared for immunoprecipitation (IP) and analyzed by western blot (WB). Bottom: Quantification of IP. Relative levels of PKM2, ENO1, PKM1, or PCK2 in IP were respectively normalized to that in WCL for each group. ( B – D ) PGAM1 directly interacts with PKM2 but not ENO1. Recombinant His-PGAM1 was immobilized on Ni-beads and then mixed with recombinant GST-PKM2 ( B ) or GST-ENO1 ( C ) prior to PD assay. The protein complex on the Ni-beads was eluted and analyzed by WB. ( D ) Quantification of PD in ( B , C ). Relative levels of GST-PKM2 or GST-ENO1 were normalized to that of His (PGAM1) for each group. ( E ) Confirmation of PGAM1 and PKM2 interaction by BLI assay. Recombinant His-PGAM1 protein was loaded onto Ni-sensor. The sensorgrams for the interaction of PGAM1 with GST-PKM2 as well as the associated KD values are shown as indicated. ( F ) PGAM1–PKM2 interaction and PGAM1 phosphorylation are more evident in tumor cells. Left: PGAM1-associated proteins were immunoprecipitated from the WCL of different tumor cells and analyzed by WB. Right: Quantification of IP. Relative levels of PKM2 or 3-pHis were normalized to that of PGAM1 for each group. ( G ) PKM2 K270M mutation reduces PGAM1 H11 phosphorylation. Left: PKM2 WT or K270M mutants were restored in endogenous PKM2-depleted A549 cells, PGAM1-associated proteins were immunoprecipitated and analyzed by WB. Right: Quantification of IP. Relative levels of PKM2 or 3-pHis were normalized to that of PGAM1 for each group. ( H ) PKM2 directly phosphorylates PGAM1 at H11. Top: In vitro kinase assays were carried out with recombinant GST-PKM2, His-PGAM1 as well as His-PGAM1 H11N mutant in the presence of PEP under normal (pH = 8.8, unboiled), heating (pH = 8.8, boiled) or acid condition (pH = 1, unboiled). PGAM1 H11 phosphorylation was detected by WB. Bottom: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. ( I , J ) PKM2 upregulates PGAM1 activity. The PGAM1 activity ( I ) and pyruvate generation ( J ) from in vitro kinase assay in the normal condition as in ( H ) were detected by commercially available kits. ( K ) ITC binding measurements of 2,3-BPG with PGAM1. The recombinant His-PGAM1 proteins purified bacterially were injected into calorimetric cells before 2,3-BPG injection. Theoretical titration curves as well as associated KD values were displayed. ( L ) DARTS binding assay of 2,3-BPG with PGAM1. A549 cells were transfected with Flag-PGAM1 WT, Y26F or Y92F for 48 h. Top: The whole lysates were respectively incubated with the indicated doses of 2,3-BPG respectively for 30 min at room temperature followed by pronase digestion and WB analysis. Bottom: Quantification of WB. Relative levels of Flag were normalized to tubulin without pronase for each group. ( M ) PKM2 upregulates H11 phosphorylation of wild-type PGAM1 as well as Y26F and Y92F mutations. Top: A549 cells were transfected with Flag-PGAM1 WT, Y26F or Y92F for 48 h. Flag (PGAM1)-associated proteins were immunoprecipitated and analyzed by WB. Bottom: Quantification of IP. Relative levels of 3-pHis or PKM2 were normalized to that of Flag (PGAM1) for each group. Data information: for WB in ( A – C , F – H , L , M ), BLI in ( E ) and ITC in ( K ), one representative experiment out of three is shown. For IP in ( A , F , G , M ), IgG served as a negative control. Data were represented as mean ± SD ( n = 3) with significance determined by one-way ( F – J , L , M ) or two-way ( A , D ) analysis of variance (ANOVA) test; **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05. ns nonsignificant. .

    Techniques Used: Immunoprecipitation, Western Blot, Recombinant, Mutagenesis, In Vitro, Activity Assay, Kinase Assay, Binding Assay, Purification, Injection, Titration, Transfection, Incubation, Negative Control

    ( A – C ) ENO1, PKM1 or ATP does not affect PGAM1 H11 phosphorylation. Left: In vitro kinase assay was carried out with recombinant GST-ENO1, GST-PKM2, GST-PKM1 and His-PGAM1 in the presence of PEP or ATP. PGAM1 H11 phosphorylation was detected by WB. Right: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. ( D ) PGAM1 does not interact with NME1 or NME2. Left: PGAM1-associated proteins in A549 cells were immunoprecipitated and analyzed by WB. IgG served as a negative control. Right: Quantification of IP. Relative levels of NME1 or NME2 in IP were normalized to that in WCL for each group. ( E – H ) NME1 or NME2 does not affect PGAM1 H11 phosphorylation. Left: In vitro kinase assay was carried out with recombinant GST-PKM2, His-NME1, NME2 and His-PGAM1 in the presence of PEP, ATP or GTP. PGAM1 H11 phosphorylation was detected by WB. Right: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. Data information: for WB in ( A – H ), one representative experiment out of three was shown. Data were represented as mean ± SD of three independent experiments ( A – H ) with significance determined by one-way ANOVA test ( A – C , E – H ) and Student’s t test ( D ); **** P < 0.0001, * P < 0.05, ns nonsignificant.
    Figure Legend Snippet: ( A – C ) ENO1, PKM1 or ATP does not affect PGAM1 H11 phosphorylation. Left: In vitro kinase assay was carried out with recombinant GST-ENO1, GST-PKM2, GST-PKM1 and His-PGAM1 in the presence of PEP or ATP. PGAM1 H11 phosphorylation was detected by WB. Right: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. ( D ) PGAM1 does not interact with NME1 or NME2. Left: PGAM1-associated proteins in A549 cells were immunoprecipitated and analyzed by WB. IgG served as a negative control. Right: Quantification of IP. Relative levels of NME1 or NME2 in IP were normalized to that in WCL for each group. ( E – H ) NME1 or NME2 does not affect PGAM1 H11 phosphorylation. Left: In vitro kinase assay was carried out with recombinant GST-PKM2, His-NME1, NME2 and His-PGAM1 in the presence of PEP, ATP or GTP. PGAM1 H11 phosphorylation was detected by WB. Right: Quantification of WB. Relative levels of 3-pHis were normalized to that of His (PGAM1) for each group. Data information: for WB in ( A – H ), one representative experiment out of three was shown. Data were represented as mean ± SD of three independent experiments ( A – H ) with significance determined by one-way ANOVA test ( A – C , E – H ) and Student’s t test ( D ); **** P < 0.0001, * P < 0.05, ns nonsignificant.

    Techniques Used: In Vitro, Kinase Assay, Recombinant, Immunoprecipitation, Negative Control

    enolase 1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc enolase 1
    Enolase 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti eno1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eno1
    Anti Eno1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eno1/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    anti eno1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eno1
    Fluvoxamine inhibits glycolysis of CD4 + T cells. A A heatmap shows the differentially expressed genes relevant to glycolysis between fluvoxamine- and vehicle-treated CD4 + T cells. The data were derived from 3 independent biological replicates. B CD4 + T cells were cultured with different concentration of fluvoxamine for 24 h in vitro and extracellular acidification rate (ECAR) was analyzed by an extracellular flux analyzer. C Glycolysis and glycolytic capacity in CD4 + T cells were determined with different dose of fluvoxamine treatment. D–J Real-time PCR analysis and Western blotting were performed to measure the expression of genes in the glycolysis pathway ( Hk2 , Pgk1 , <t>Eno1</t> , Glut1 , Pkm2 and Ldha ) in CD4 + T cells with/without fluvoxamine treatment. Each dot represents the mean of three biological replicates. Data are expressed as mean ± SEM. Statistical significance was calculated by unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, not significant. Statistical difference in C was analyzed by one-way ANOVA
    Anti Eno1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eno1/product/Cell Signaling Technology Inc
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    1) Product Images from "Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes"

    Article Title: Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes

    Journal: Molecular Medicine

    doi: 10.1186/s10020-024-00791-1

    Fluvoxamine inhibits glycolysis of CD4 + T cells. A A heatmap shows the differentially expressed genes relevant to glycolysis between fluvoxamine- and vehicle-treated CD4 + T cells. The data were derived from 3 independent biological replicates. B CD4 + T cells were cultured with different concentration of fluvoxamine for 24 h in vitro and extracellular acidification rate (ECAR) was analyzed by an extracellular flux analyzer. C Glycolysis and glycolytic capacity in CD4 + T cells were determined with different dose of fluvoxamine treatment. D–J Real-time PCR analysis and Western blotting were performed to measure the expression of genes in the glycolysis pathway ( Hk2 , Pgk1 , Eno1 , Glut1 , Pkm2 and Ldha ) in CD4 + T cells with/without fluvoxamine treatment. Each dot represents the mean of three biological replicates. Data are expressed as mean ± SEM. Statistical significance was calculated by unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, not significant. Statistical difference in C was analyzed by one-way ANOVA
    Figure Legend Snippet: Fluvoxamine inhibits glycolysis of CD4 + T cells. A A heatmap shows the differentially expressed genes relevant to glycolysis between fluvoxamine- and vehicle-treated CD4 + T cells. The data were derived from 3 independent biological replicates. B CD4 + T cells were cultured with different concentration of fluvoxamine for 24 h in vitro and extracellular acidification rate (ECAR) was analyzed by an extracellular flux analyzer. C Glycolysis and glycolytic capacity in CD4 + T cells were determined with different dose of fluvoxamine treatment. D–J Real-time PCR analysis and Western blotting were performed to measure the expression of genes in the glycolysis pathway ( Hk2 , Pgk1 , Eno1 , Glut1 , Pkm2 and Ldha ) in CD4 + T cells with/without fluvoxamine treatment. Each dot represents the mean of three biological replicates. Data are expressed as mean ± SEM. Statistical significance was calculated by unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, not significant. Statistical difference in C was analyzed by one-way ANOVA

    Techniques Used: Derivative Assay, Cell Culture, Concentration Assay, In Vitro, Real-time Polymerase Chain Reaction, Western Blot, Expressing

    anti eno1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eno1
    Anti Eno1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    eno1 3810s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc eno1 3810s
    Eno1 3810s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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