anti eno1 antibodies Search Results


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  • 91
    StressMarq inhibitory site tyr44
    Inhibitory Site Tyr44, supplied by StressMarq, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/inhibitory site tyr44/product/StressMarq
    Average 91 stars, based on 1 article reviews
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    inhibitory site tyr44 - by Bioz Stars, 2024-06
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    86
    Proteintech anti eno1 antibody
    Reduced CAA pathology and improved cognitive impairment in APP23 mice by <t>ENO1.</t> a Schedule of administration of <t>ENO1</t> (or vehicle control) and behavioral testing for 18-month-old male APP23 and wild-type mice. ENO1 (1 mM, 100 μl) or vehicle control (100 μl) was infused into the brain continuously for 7 days. b Representative immunoblot of 18-month-old male APP23 mouse brain lysates treated with ENO1 (right) and vehicle control (left), analyzed by using anti-human Aβ antibody (6E10), with quantification of relative intensity of monomeric Aβ. c Representative dot blots of 18-month-old male APP23 mouse brain lysates treated with ENO1 (right) and vehicle control (left), analyzed using anti-Aβ fibril antibody, with quantification of relative intensity of Aβ fibrils. d–g Quantification of soluble and insoluble Aβ40 and Aβ42 (n = 3 per group). h Representative immunoblots of lysates from the hippocampus (left) and cerebral cortex (right) showing APP expression, with quantification of the relative intensities of APP (n = 3 per group). i, j Y-maze test results. Average alternation (%) (i) and total number of entries into each arm (j) for each group (vehicle-infused wild-type mice, n = 6; ENO1-infused wild-type mice, n = 5; vehicle-infused APP23 mice, n = 7; ENO1-infused APP23 mice, n = 7, heat-inactivated ENO1-infused APP23 mice, n = 5). k Representative histopathological images of cerebrovascular Aβ deposits (upper) and parenchymal Aβ plaques (lower) in vehicle-infused, ENO1-infused, and heat-inactivated ENO1-infused APP23 mice. Scale bars = 500 μm (upper), 100 μm (lower). l, m Quantitative analyses of the numbers of vessels with Aβ deposits (l), as well as the percent area of Aβ plaques (m) (vehicle-infused APP23, n = 4; ENO1-infused APP23, n = 4; heat-inactivated ENO1-infused APP23, n = 5). Monomeric Aβ, Aβ fibrils, and APP protein levels were normalized to β-actin protein levels. Data are shown as mean ± SEM. *P < 0.05, ***P < 0.001. NS not significant
    Anti Eno1 Antibody, supplied by Proteintech, 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 antibody/product/Proteintech
    Average 86 stars, based on 1 article reviews
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    anti eno1 antibody - by Bioz Stars, 2024-06
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    97
    Santa Cruz Biotechnology eno1
    Identified proteins altered by age and E 2
    Eno1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Abcam anti eno1 antibody
    Protein Identification Summary.
    Anti Eno1 Antibody, supplied by Abcam, 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 antibody/product/Abcam
    Average 86 stars, based on 1 article reviews
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    Image Search Results


    Reduced CAA pathology and improved cognitive impairment in APP23 mice by ENO1. a Schedule of administration of ENO1 (or vehicle control) and behavioral testing for 18-month-old male APP23 and wild-type mice. ENO1 (1 mM, 100 μl) or vehicle control (100 μl) was infused into the brain continuously for 7 days. b Representative immunoblot of 18-month-old male APP23 mouse brain lysates treated with ENO1 (right) and vehicle control (left), analyzed by using anti-human Aβ antibody (6E10), with quantification of relative intensity of monomeric Aβ. c Representative dot blots of 18-month-old male APP23 mouse brain lysates treated with ENO1 (right) and vehicle control (left), analyzed using anti-Aβ fibril antibody, with quantification of relative intensity of Aβ fibrils. d–g Quantification of soluble and insoluble Aβ40 and Aβ42 (n = 3 per group). h Representative immunoblots of lysates from the hippocampus (left) and cerebral cortex (right) showing APP expression, with quantification of the relative intensities of APP (n = 3 per group). i, j Y-maze test results. Average alternation (%) (i) and total number of entries into each arm (j) for each group (vehicle-infused wild-type mice, n = 6; ENO1-infused wild-type mice, n = 5; vehicle-infused APP23 mice, n = 7; ENO1-infused APP23 mice, n = 7, heat-inactivated ENO1-infused APP23 mice, n = 5). k Representative histopathological images of cerebrovascular Aβ deposits (upper) and parenchymal Aβ plaques (lower) in vehicle-infused, ENO1-infused, and heat-inactivated ENO1-infused APP23 mice. Scale bars = 500 μm (upper), 100 μm (lower). l, m Quantitative analyses of the numbers of vessels with Aβ deposits (l), as well as the percent area of Aβ plaques (m) (vehicle-infused APP23, n = 4; ENO1-infused APP23, n = 4; heat-inactivated ENO1-infused APP23, n = 5). Monomeric Aβ, Aβ fibrils, and APP protein levels were normalized to β-actin protein levels. Data are shown as mean ± SEM. *P < 0.05, ***P < 0.001. NS not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Reduced CAA pathology and improved cognitive impairment in APP23 mice by ENO1. a Schedule of administration of ENO1 (or vehicle control) and behavioral testing for 18-month-old male APP23 and wild-type mice. ENO1 (1 mM, 100 μl) or vehicle control (100 μl) was infused into the brain continuously for 7 days. b Representative immunoblot of 18-month-old male APP23 mouse brain lysates treated with ENO1 (right) and vehicle control (left), analyzed by using anti-human Aβ antibody (6E10), with quantification of relative intensity of monomeric Aβ. c Representative dot blots of 18-month-old male APP23 mouse brain lysates treated with ENO1 (right) and vehicle control (left), analyzed using anti-Aβ fibril antibody, with quantification of relative intensity of Aβ fibrils. d–g Quantification of soluble and insoluble Aβ40 and Aβ42 (n = 3 per group). h Representative immunoblots of lysates from the hippocampus (left) and cerebral cortex (right) showing APP expression, with quantification of the relative intensities of APP (n = 3 per group). i, j Y-maze test results. Average alternation (%) (i) and total number of entries into each arm (j) for each group (vehicle-infused wild-type mice, n = 6; ENO1-infused wild-type mice, n = 5; vehicle-infused APP23 mice, n = 7; ENO1-infused APP23 mice, n = 7, heat-inactivated ENO1-infused APP23 mice, n = 5). k Representative histopathological images of cerebrovascular Aβ deposits (upper) and parenchymal Aβ plaques (lower) in vehicle-infused, ENO1-infused, and heat-inactivated ENO1-infused APP23 mice. Scale bars = 500 μm (upper), 100 μm (lower). l, m Quantitative analyses of the numbers of vessels with Aβ deposits (l), as well as the percent area of Aβ plaques (m) (vehicle-infused APP23, n = 4; ENO1-infused APP23, n = 4; heat-inactivated ENO1-infused APP23, n = 5). Monomeric Aβ, Aβ fibrils, and APP protein levels were normalized to β-actin protein levels. Data are shown as mean ± SEM. *P < 0.05, ***P < 0.001. NS not significant

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Western Blot, Expressing

    Interaction of Aβ and ENO1 in vitro. a, b ThT fluorescence of Aβ—Aβ40, 50 μM (a); Aβ42, 10 μM (b)—incubated with ENO1 at a final concentration of 1 μM. c, d Representative TEM images of Aβ40 incubated with ENO1 (d) and Aβ42 incubated with ENO1 (d). Scale bars = 2 μm. e, f Representative immunoblots of Aβ40 incubated with ENO1 (e) and Aβ42 incubated with ENO1 (f), analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. g, h ThT fluorescence of Aβ fibrils—Aβ40 fibrils, 50 μM (g); Aβ42 fibrils, 10 μM (h)—incubated with ENO1 at a final concentration of 1 μM. i, j Representative TEM images of Aβ40 fibrils incubated with ENO1 (i) and Aβ42 fibrils incubated with ENO1 (j). Scale bars = 2 μm. k, l Representative immunoblots of Aβ40 fibrils incubated with ENO1 (k) and Aβ42 fibrils incubated with ENO1 (l), analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. Data are shown as mean ± SEM of three independent experiments. ***P < 0.001. NS not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Interaction of Aβ and ENO1 in vitro. a, b ThT fluorescence of Aβ—Aβ40, 50 μM (a); Aβ42, 10 μM (b)—incubated with ENO1 at a final concentration of 1 μM. c, d Representative TEM images of Aβ40 incubated with ENO1 (d) and Aβ42 incubated with ENO1 (d). Scale bars = 2 μm. e, f Representative immunoblots of Aβ40 incubated with ENO1 (e) and Aβ42 incubated with ENO1 (f), analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. g, h ThT fluorescence of Aβ fibrils—Aβ40 fibrils, 50 μM (g); Aβ42 fibrils, 10 μM (h)—incubated with ENO1 at a final concentration of 1 μM. i, j Representative TEM images of Aβ40 fibrils incubated with ENO1 (i) and Aβ42 fibrils incubated with ENO1 (j). Scale bars = 2 μm. k, l Representative immunoblots of Aβ40 fibrils incubated with ENO1 (k) and Aβ42 fibrils incubated with ENO1 (l), analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. Data are shown as mean ± SEM of three independent experiments. ***P < 0.001. NS not significant

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: In Vitro, Fluorescence, Incubation, Concentration Assay, Western Blot

    Reversal of the inhibitory effects of ENO1 on Aβ fibril formation by the ENO1 inhibitor NaF. a ThT fluorescence kinetics of Aβ40 (50 μM) incubated with ENO1 (1 μM) and NaF at the indicated concentrations (10 mM, 1 mM, and 100 μM) for 7 days. b Representative TEM images of Aβ40 incubated with ENO1 and NaF. Scale bars = 2 μm. c Representative immunoblot of Aβ40 incubated with ENO1 and NaF, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. d ThT fluorescence kinetics of Aβ42 (10 μM) incubated with ENO1 (1 μM) and NaF at the indicated concentrations for 7 days. e Representative TEM images of Aβ42 incubated with ENO1 and NaF. Scale bars = 2 μm. f Representative immunoblot of Aβ42 incubated with ENO1 and NaF, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative densities of Aβ fibrils. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Reversal of the inhibitory effects of ENO1 on Aβ fibril formation by the ENO1 inhibitor NaF. a ThT fluorescence kinetics of Aβ40 (50 μM) incubated with ENO1 (1 μM) and NaF at the indicated concentrations (10 mM, 1 mM, and 100 μM) for 7 days. b Representative TEM images of Aβ40 incubated with ENO1 and NaF. Scale bars = 2 μm. c Representative immunoblot of Aβ40 incubated with ENO1 and NaF, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. d ThT fluorescence kinetics of Aβ42 (10 μM) incubated with ENO1 (1 μM) and NaF at the indicated concentrations for 7 days. e Representative TEM images of Aβ42 incubated with ENO1 and NaF. Scale bars = 2 μm. f Representative immunoblot of Aβ42 incubated with ENO1 and NaF, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative densities of Aβ fibrils. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS not significant

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Fluorescence, Incubation, Western Blot

    Induction of ENO1 expression by Aβ. a, b ENO1 mRNA levels in cerebrovascular smooth muscle cells treated with freshly prepared Aβ40 (a) or Aβ42 (b) at a final concentration of 10 μM for 24 h, or treated with PBS (control). ENO1 mRNA levels were normalized to β-actin mRNA levels. c, d Representative immunoblots and quantification of the relative ENO1 protein levels of cultured cell lysates treated with freshly prepared Aβ40 (c) or Aβ42 (d) at a final concentration of 10 μM for 24 h, or treated with PBS (control). e, f Representative immunoblots and quantification of the relative ENO1 protein levels in culture medium treated with freshly prepared Aβ40 (e) or Aβ42 (f). g, h ENO1 mRNA levels in cerebrovascular smooth muscle cells treated with Aβ40 fibrils (g) or Aβ42 fibrils (h) at a final concentration of 10 μM for 24 h, or treated with PBS (control). ENO1 mRNA levels were normalized to β-actin mRNA levels. i, j Representative immunoblots and quantification of the relative ENO1 protein levels of cultured cell lysates treated with Aβ40 fibrils (i) or Aβ42 fibrils (j) at a final concentration of 10 μM for 24 h, or treated with PBS (control). k, l Representative immunoblots and quantification of the relative ENO1 protein levels in culture medium treated with Aβ40 fibrils (k) or Aβ42 fibrils (l). m ENO1 mRNA levels in lysates of cultured murine cerebrovascular smooth muscle cells treated with H2O2 for 24 h. n, o Representative immunofluorescence images—ENO1 (green) and Aβ (red)—of cerebrovascular Aβ deposits (n) and parenchymal Aβ deposits (o) in 18-month-old APP23 mice. p, q ENO1 mRNA levels in the hippocampus, cerebral cortex, and cerebral blood vessels from 4-month-old wild type (n = 6) and APP23 (n = 4) mice (p), and from 18-month-old wild type (n = 7) and APP23 (n = 5) mice (q). ENO1 mRNA and protein levels were normalized to β-actin mRNA and protein levels. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus control. NS not significant. Scale bars = 50 μm

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Induction of ENO1 expression by Aβ. a, b ENO1 mRNA levels in cerebrovascular smooth muscle cells treated with freshly prepared Aβ40 (a) or Aβ42 (b) at a final concentration of 10 μM for 24 h, or treated with PBS (control). ENO1 mRNA levels were normalized to β-actin mRNA levels. c, d Representative immunoblots and quantification of the relative ENO1 protein levels of cultured cell lysates treated with freshly prepared Aβ40 (c) or Aβ42 (d) at a final concentration of 10 μM for 24 h, or treated with PBS (control). e, f Representative immunoblots and quantification of the relative ENO1 protein levels in culture medium treated with freshly prepared Aβ40 (e) or Aβ42 (f). g, h ENO1 mRNA levels in cerebrovascular smooth muscle cells treated with Aβ40 fibrils (g) or Aβ42 fibrils (h) at a final concentration of 10 μM for 24 h, or treated with PBS (control). ENO1 mRNA levels were normalized to β-actin mRNA levels. i, j Representative immunoblots and quantification of the relative ENO1 protein levels of cultured cell lysates treated with Aβ40 fibrils (i) or Aβ42 fibrils (j) at a final concentration of 10 μM for 24 h, or treated with PBS (control). k, l Representative immunoblots and quantification of the relative ENO1 protein levels in culture medium treated with Aβ40 fibrils (k) or Aβ42 fibrils (l). m ENO1 mRNA levels in lysates of cultured murine cerebrovascular smooth muscle cells treated with H2O2 for 24 h. n, o Representative immunofluorescence images—ENO1 (green) and Aβ (red)—of cerebrovascular Aβ deposits (n) and parenchymal Aβ deposits (o) in 18-month-old APP23 mice. p, q ENO1 mRNA levels in the hippocampus, cerebral cortex, and cerebral blood vessels from 4-month-old wild type (n = 6) and APP23 (n = 4) mice (p), and from 18-month-old wild type (n = 7) and APP23 (n = 5) mice (q). ENO1 mRNA and protein levels were normalized to β-actin mRNA and protein levels. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus control. NS not significant. Scale bars = 50 μm

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Expressing, Concentration Assay, Western Blot, Cell Culture, Immunofluorescence

    ENO1-induced reduced Aβ cytotoxicity in cerebrovascular smooth muscle cells. a Significantly reduced Aβ40-induced caspase-3/7 activity after administration of ENO1 together with Aβ40 for 24 h. b Representative images from immunofluorescence studies with anti-human Aβ antibody (1:100 dilution, 6E10) in cultured cells treated with Aβ40 (10 μM) for 24 h in the absence (left) or presence (right) of ENO1 (1 μM). Scale bars = 100 μm. c Quantification of the percentages of Aβ40-positive areas obtained by averaging 10 random fields of cultured cells in the absence or presence of ENO1. d Enhancement of Aβ40-induced caspase-3/7 activity by ENO1 knockdown. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, ***P < 0.001. NS not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: ENO1-induced reduced Aβ cytotoxicity in cerebrovascular smooth muscle cells. a Significantly reduced Aβ40-induced caspase-3/7 activity after administration of ENO1 together with Aβ40 for 24 h. b Representative images from immunofluorescence studies with anti-human Aβ antibody (1:100 dilution, 6E10) in cultured cells treated with Aβ40 (10 μM) for 24 h in the absence (left) or presence (right) of ENO1 (1 μM). Scale bars = 100 μm. c Quantification of the percentages of Aβ40-positive areas obtained by averaging 10 random fields of cultured cells in the absence or presence of ENO1. d Enhancement of Aβ40-induced caspase-3/7 activity by ENO1 knockdown. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, ***P < 0.001. NS not significant

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Activity Assay, Immunofluorescence, Cell Culture

    Reversal of the inhibitory effects of ENO1 on Aβ fibril formation by Pefabloc, a serine protease inhibitor. a ThT fluorescence kinetics of Aβ40 (50 μM) incubated with ENO1 (1 μM) and Pefabloc (pefa) for 7 days. b Representative TEM images of Aβ40 incubated with ENO1 and Pefabloc. Scale bars = 2 μm. c Representative immunoblot of Aβ40 incubated with ENO1 and Pefabloc, analyzed by using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. d Total amount of uncleaved Aβ (left), and total amount of cleaved Aβ (right), with quantification of the relative densities of Aβ. e–h ThT fluorescence kinetics of Aβ40 (50 μM) incubated with ENO1 (1 μM) and protease inhibitors (described below) for 7 days. i-l Representative TEM images of Aβ40 incubated with ENO1 and protease inhibitors. Scale bars = 2 μm. m-p Representative immunoblots of Aβ40 incubated with ENO1 and protease inhibitors, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. Protease inhibitors were used at the indicated concentrations: Pefabloc SC, a serine protease inhibitor, 2.5 mM, 1 mM; pepstatin A, an aspartic protease inhibitor, 10 μM, 1 μM; E-64, a cysteine protease inhibitor, 1 mM, 100 μM; phosphoramidon, a metalloprotease inhibitor, 1 mM, 100 μM; and EDTA, a metalloprotease inhibitor, 5 mM, 1 mM. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, ***P < 0.001. NS not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Reversal of the inhibitory effects of ENO1 on Aβ fibril formation by Pefabloc, a serine protease inhibitor. a ThT fluorescence kinetics of Aβ40 (50 μM) incubated with ENO1 (1 μM) and Pefabloc (pefa) for 7 days. b Representative TEM images of Aβ40 incubated with ENO1 and Pefabloc. Scale bars = 2 μm. c Representative immunoblot of Aβ40 incubated with ENO1 and Pefabloc, analyzed by using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. d Total amount of uncleaved Aβ (left), and total amount of cleaved Aβ (right), with quantification of the relative densities of Aβ. e–h ThT fluorescence kinetics of Aβ40 (50 μM) incubated with ENO1 (1 μM) and protease inhibitors (described below) for 7 days. i-l Representative TEM images of Aβ40 incubated with ENO1 and protease inhibitors. Scale bars = 2 μm. m-p Representative immunoblots of Aβ40 incubated with ENO1 and protease inhibitors, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensities of Aβ fibrils. Protease inhibitors were used at the indicated concentrations: Pefabloc SC, a serine protease inhibitor, 2.5 mM, 1 mM; pepstatin A, an aspartic protease inhibitor, 10 μM, 1 μM; E-64, a cysteine protease inhibitor, 1 mM, 100 μM; phosphoramidon, a metalloprotease inhibitor, 1 mM, 100 μM; and EDTA, a metalloprotease inhibitor, 5 mM, 1 mM. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, ***P < 0.001. NS not significant

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Protease Inhibitor, Fluorescence, Incubation, Western Blot

    Nitrative modification of ENO1 in vivo and in vitro. a Representative immunoblots of immunoprecipitated ENO1 from the hippocampus, cerebral cortex, and cerebral blood vessels obtained from 18-month-old wild-type and APP23 mice; probes were anti-3-nitrotyrosine and anti-ENO1 antibodies. Quantified nitrated ENO1 levels in the hippocampus, cerebral cortex, and cerebral blood vessels appear below the immunoblots. Data were normalized to ENO1 protein levels. b Representative immunoblots of ENO1, analyzed using anti-3-nitrotyrosine and anti-ENO1 antibodies, with quantification of the relative intensity of nitrated ENO1. Data were normalized to ENO1 protein levels. c Quantified ENO1 enzymatic activity of non-nitrated ENO1 and nitrated ENO1. d, e Representative immunofluorescence images—ENO1 (green) and 3-nitrotyrosine (red)—of cerebrovascular Aβ deposits (d) and parenchymal Aβ deposits (e) in 18-month-old APP23 mice. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS not significant. Scale bars = 50 μm

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Nitrative modification of ENO1 in vivo and in vitro. a Representative immunoblots of immunoprecipitated ENO1 from the hippocampus, cerebral cortex, and cerebral blood vessels obtained from 18-month-old wild-type and APP23 mice; probes were anti-3-nitrotyrosine and anti-ENO1 antibodies. Quantified nitrated ENO1 levels in the hippocampus, cerebral cortex, and cerebral blood vessels appear below the immunoblots. Data were normalized to ENO1 protein levels. b Representative immunoblots of ENO1, analyzed using anti-3-nitrotyrosine and anti-ENO1 antibodies, with quantification of the relative intensity of nitrated ENO1. Data were normalized to ENO1 protein levels. c Quantified ENO1 enzymatic activity of non-nitrated ENO1 and nitrated ENO1. d, e Representative immunofluorescence images—ENO1 (green) and 3-nitrotyrosine (red)—of cerebrovascular Aβ deposits (d) and parenchymal Aβ deposits (e) in 18-month-old APP23 mice. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS not significant. Scale bars = 50 μm

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Modification, In Vivo, In Vitro, Western Blot, Immunoprecipitation, Activity Assay, Immunofluorescence

    Elimination by heat-inactivated ENO1 of the inhibitory effects of ENO1 on Aβ40 fibril formation. a Quantified enzymatic activities of ENO1. b ThT fluorescence kinetics of Aβ40 (50 μM) incubated with heat-treated ENO1 (60 ℃ or 90 ℃ for 5 min) at a final concentration of 1 μM for 7 days. c Representative TEM images of Aβ40 incubated alone, with ENO1, and with heat-treated ENO1. Scale bars = 2 μm. d Representative immunoblot of Aβ40 incubated alone, with ENO1, and with heat-treated ENO1, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensity of Aβ40 fibrils. e The bar graph shows ThT fluorescence values on Day 3; Aβ40 fibrils (50 μM) incubated with heat-treated ENO1 (90 ℃ for 5 min) at a final concentration of 1 μM for 3 days. f Representative TEM images Aβ40 fibrils incubated alone, or incubated with heat-treated ENO1 (90 ℃ for 5 min). Scale bars = 2 μm. g Representative immunoblot of Aβ40 fibrils incubated alone, or incubated with heat-treated ENO1 (90 ℃ for 5 min), analyzed using anti-human Aβ antibody (6E10), with quantification of relative intensity of Aβ40 fibrils. h ThT fluorescence kinetics of Aβ42 (10 μM) incubated with heat-treated ENO1 at a final concentration of 1 μM for 7 days. i Representative TEM images of Aβ42 incubated alone, with ENO1, and with heat-treated ENO1 (60 ℃ or 90 ℃ for 5 min). Scale bars = 2 μm. j Representative immunoblot of Aβ42 incubated with heat-treated ENO1, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative densities of Aβ42 fibrils. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: α-Enolase reduces cerebrovascular Aβ deposits by protecting Aβ amyloid formation

    doi: 10.1007/s00018-022-04493-x

    Figure Lengend Snippet: Elimination by heat-inactivated ENO1 of the inhibitory effects of ENO1 on Aβ40 fibril formation. a Quantified enzymatic activities of ENO1. b ThT fluorescence kinetics of Aβ40 (50 μM) incubated with heat-treated ENO1 (60 ℃ or 90 ℃ for 5 min) at a final concentration of 1 μM for 7 days. c Representative TEM images of Aβ40 incubated alone, with ENO1, and with heat-treated ENO1. Scale bars = 2 μm. d Representative immunoblot of Aβ40 incubated alone, with ENO1, and with heat-treated ENO1, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative intensity of Aβ40 fibrils. e The bar graph shows ThT fluorescence values on Day 3; Aβ40 fibrils (50 μM) incubated with heat-treated ENO1 (90 ℃ for 5 min) at a final concentration of 1 μM for 3 days. f Representative TEM images Aβ40 fibrils incubated alone, or incubated with heat-treated ENO1 (90 ℃ for 5 min). Scale bars = 2 μm. g Representative immunoblot of Aβ40 fibrils incubated alone, or incubated with heat-treated ENO1 (90 ℃ for 5 min), analyzed using anti-human Aβ antibody (6E10), with quantification of relative intensity of Aβ40 fibrils. h ThT fluorescence kinetics of Aβ42 (10 μM) incubated with heat-treated ENO1 at a final concentration of 1 μM for 7 days. i Representative TEM images of Aβ42 incubated alone, with ENO1, and with heat-treated ENO1 (60 ℃ or 90 ℃ for 5 min). Scale bars = 2 μm. j Representative immunoblot of Aβ42 incubated with heat-treated ENO1, analyzed using anti-human Aβ antibody (6E10), with quantification of the relative densities of Aβ42 fibrils. Data are shown as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS not significant

    Article Snippet: We then used Western blotting to quantify nitrated ENO1 levels using an anti-ENO1 antibody (1:1000 dilution; Proteintech Group) and an anti-3-nitrotyrosine antibody (1:1000 dilution; Abcam).

    Techniques: Fluorescence, Incubation, Concentration Assay, Western Blot

    Identified proteins altered by age and E 2

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Age-dependent Effects of 17β-estradiol on the Dynamics of Estrogen Receptor β (ERβ) Protein–Protein Interactions in the Ventral Hippocampus

    doi: 10.1074/mcp.M113.031559

    Figure Lengend Snippet: Identified proteins altered by age and E 2

    Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) for 1 h before the addition of 1° antibody in TBST with 5% BSA and 0.01% NaN 3 for 1.5 h. All 1° antibodies were used at a 1:1000 dilution: VCP (Pierce, PA5–17486), ERβ (Santa Cruz, Sc-8974x), ENO1 (Santa Cruz, sc-15343), GAPDH (Santa Cruz, sc-25778), GELS (Cell Signaling, #8090), HnRNPH (Santa Cruz, sc-15387), HSP70 (GenTex, GTX-104126), and β-actin (Cell Signaling, Aurora, IL, 4970S).

    Techniques: Molecular Weight, Binding Assay

    DeCyder topography, gel image analysis, and average log standard abundance of α-enolase (ENO1) in response to E2 in young and aged animals. For each panel from top left to right, 3 month: YV representative topography, YE representative topography, YV representative gel image, and YE representative gel image. 18 month: AV representative topography, AE representative topography, AV representative gel image, and AE representative gel image. Graph depicts log transformed average abundance normalized to internal standard and matched to master gel. Average calculated from three independent experiments with a biological variance of four pooled animals per experiment (n = 3, BV = 12). *Significance from vehicle, p < 0.05.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Age-dependent Effects of 17β-estradiol on the Dynamics of Estrogen Receptor β (ERβ) Protein–Protein Interactions in the Ventral Hippocampus

    doi: 10.1074/mcp.M113.031559

    Figure Lengend Snippet: DeCyder topography, gel image analysis, and average log standard abundance of α-enolase (ENO1) in response to E2 in young and aged animals. For each panel from top left to right, 3 month: YV representative topography, YE representative topography, YV representative gel image, and YE representative gel image. 18 month: AV representative topography, AE representative topography, AV representative gel image, and AE representative gel image. Graph depicts log transformed average abundance normalized to internal standard and matched to master gel. Average calculated from three independent experiments with a biological variance of four pooled animals per experiment (n = 3, BV = 12). *Significance from vehicle, p < 0.05.

    Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) for 1 h before the addition of 1° antibody in TBST with 5% BSA and 0.01% NaN 3 for 1.5 h. All 1° antibodies were used at a 1:1000 dilution: VCP (Pierce, PA5–17486), ERβ (Santa Cruz, Sc-8974x), ENO1 (Santa Cruz, sc-15343), GAPDH (Santa Cruz, sc-25778), GELS (Cell Signaling, #8090), HnRNPH (Santa Cruz, sc-15387), HSP70 (GenTex, GTX-104126), and β-actin (Cell Signaling, Aurora, IL, 4970S).

    Techniques: Transformation Assay

    Confirmation of ERβ–VCP interaction and subcellular expression of ERβ-interaction partners. A, confirmation of ERβ–VCP interaction in ventral hippocampus by immunoblot with corresponding input and nonspecific IgG control. B, representative immunoblots for nuclear and cytosolic ERβ, HSP70, GAPDH, VCP, HNRNP H, and ENO1 expression in ventral hippocampus normalized to β-actin. C, quantification of densitometric analysis of protein expression calculated from at least three independent experiments (n = 3). *Significant difference between groups (two-way ANOVA, p < 0.05).

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Age-dependent Effects of 17β-estradiol on the Dynamics of Estrogen Receptor β (ERβ) Protein–Protein Interactions in the Ventral Hippocampus

    doi: 10.1074/mcp.M113.031559

    Figure Lengend Snippet: Confirmation of ERβ–VCP interaction and subcellular expression of ERβ-interaction partners. A, confirmation of ERβ–VCP interaction in ventral hippocampus by immunoblot with corresponding input and nonspecific IgG control. B, representative immunoblots for nuclear and cytosolic ERβ, HSP70, GAPDH, VCP, HNRNP H, and ENO1 expression in ventral hippocampus normalized to β-actin. C, quantification of densitometric analysis of protein expression calculated from at least three independent experiments (n = 3). *Significant difference between groups (two-way ANOVA, p < 0.05).

    Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) for 1 h before the addition of 1° antibody in TBST with 5% BSA and 0.01% NaN 3 for 1.5 h. All 1° antibodies were used at a 1:1000 dilution: VCP (Pierce, PA5–17486), ERβ (Santa Cruz, Sc-8974x), ENO1 (Santa Cruz, sc-15343), GAPDH (Santa Cruz, sc-25778), GELS (Cell Signaling, #8090), HnRNPH (Santa Cruz, sc-15387), HSP70 (GenTex, GTX-104126), and β-actin (Cell Signaling, Aurora, IL, 4970S).

    Techniques: Expressing, Western Blot

    Protein Identification Summary.

    Journal: PLoS ONE

    Article Title: Proteomic Profiling of SupT1 Cells Reveal Modulation of Host Proteins by HIV-1 Nef Variants

    doi: 10.1371/journal.pone.0122994

    Figure Lengend Snippet: Protein Identification Summary.

    Article Snippet: Passive Lysis Buffer (Promega), PVDF membrane (Millipore), Luminata forte Western HRP substrate (Millipore), anti alpha-tubulin monoclonal antibody (Invitrogen Life Technologies), DAPI (Invitrogen Life Technologies), Primary antibodies- anti-Cyclophilin A antibody (ab58144), anti-eIF5A antibody [EP527Y] (ab32407), anti-ENO1 antibody (ab85086), anti-OTUB1 antibody (ab98280), anti-RhoGDI antibody [1F2] (ab118159), and anti-VDAC1/Porin antibody—N-terminal (ab135585) were purchased from abcam, UK.

    Techniques: Sequencing, Binding Assay, Activity Assay, Translocation Assay, Transduction, Activation Assay, Conjugation Assay

    Western blotting of representative proteins revealed similar quantitative profiles as suggested by 2DGE analysis. Contrasting effect of Nef variants observed upon expression of the six proteins- Cyclophilin A, EIF5A-1, Rho GDI, VDAC1, OTUB1, and ENO1. alpha tubulin was used as a loading control. Data are presented as the mean ± SD of three independent experiment.

    Journal: PLoS ONE

    Article Title: Proteomic Profiling of SupT1 Cells Reveal Modulation of Host Proteins by HIV-1 Nef Variants

    doi: 10.1371/journal.pone.0122994

    Figure Lengend Snippet: Western blotting of representative proteins revealed similar quantitative profiles as suggested by 2DGE analysis. Contrasting effect of Nef variants observed upon expression of the six proteins- Cyclophilin A, EIF5A-1, Rho GDI, VDAC1, OTUB1, and ENO1. alpha tubulin was used as a loading control. Data are presented as the mean ± SD of three independent experiment.

    Article Snippet: Passive Lysis Buffer (Promega), PVDF membrane (Millipore), Luminata forte Western HRP substrate (Millipore), anti alpha-tubulin monoclonal antibody (Invitrogen Life Technologies), DAPI (Invitrogen Life Technologies), Primary antibodies- anti-Cyclophilin A antibody (ab58144), anti-eIF5A antibody [EP527Y] (ab32407), anti-ENO1 antibody (ab85086), anti-OTUB1 antibody (ab98280), anti-RhoGDI antibody [1F2] (ab118159), and anti-VDAC1/Porin antibody—N-terminal (ab135585) were purchased from abcam, UK.

    Techniques: Western Blot, Expressing

    Immunofluorescence study to compare cell surface expression of six proteins in SupT1 cells (1 μm section) was captured by Confocal microscopy at 40 X and 63 X magnification. Figure shows immuostained SupT1 cells for expression of Vector/Nef (green), respective proteins (red) with their nucleus stained with DAPI (blue). Fluorescence intensity was measured for 5–8 Nef/vector transfected cells from 6 different fields of each sample and mean was calculated. Data is presented as values from two different experiments. displays ENO1 and VDAC1 (A-C) and EIF5A, OTUB1, CYPA and RhoGDI (D-H).

    Journal: PLoS ONE

    Article Title: Proteomic Profiling of SupT1 Cells Reveal Modulation of Host Proteins by HIV-1 Nef Variants

    doi: 10.1371/journal.pone.0122994

    Figure Lengend Snippet: Immunofluorescence study to compare cell surface expression of six proteins in SupT1 cells (1 μm section) was captured by Confocal microscopy at 40 X and 63 X magnification. Figure shows immuostained SupT1 cells for expression of Vector/Nef (green), respective proteins (red) with their nucleus stained with DAPI (blue). Fluorescence intensity was measured for 5–8 Nef/vector transfected cells from 6 different fields of each sample and mean was calculated. Data is presented as values from two different experiments. displays ENO1 and VDAC1 (A-C) and EIF5A, OTUB1, CYPA and RhoGDI (D-H).

    Article Snippet: Passive Lysis Buffer (Promega), PVDF membrane (Millipore), Luminata forte Western HRP substrate (Millipore), anti alpha-tubulin monoclonal antibody (Invitrogen Life Technologies), DAPI (Invitrogen Life Technologies), Primary antibodies- anti-Cyclophilin A antibody (ab58144), anti-eIF5A antibody [EP527Y] (ab32407), anti-ENO1 antibody (ab85086), anti-OTUB1 antibody (ab98280), anti-RhoGDI antibody [1F2] (ab118159), and anti-VDAC1/Porin antibody—N-terminal (ab135585) were purchased from abcam, UK.

    Techniques: Immunofluorescence, Expressing, Confocal Microscopy, Plasmid Preparation, Staining, Fluorescence, Transfection

    (A)Western blots showing reverse effect of Nef RP01 mutant upon downmodulation of α-enolase (ENO1) and VDAC1 as caused by Nef RP01. Alpha-tubulin was used as a loading control. (B)Graphs representing the difference in effect of Nef RP14 Nef RP01 and Nef RP01 mutant upon expression of α-enolase (ENO1) and VDAC1. Data are presented as the mean ± SD of two independent experiment.

    Journal: PLoS ONE

    Article Title: Proteomic Profiling of SupT1 Cells Reveal Modulation of Host Proteins by HIV-1 Nef Variants

    doi: 10.1371/journal.pone.0122994

    Figure Lengend Snippet: (A)Western blots showing reverse effect of Nef RP01 mutant upon downmodulation of α-enolase (ENO1) and VDAC1 as caused by Nef RP01. Alpha-tubulin was used as a loading control. (B)Graphs representing the difference in effect of Nef RP14 Nef RP01 and Nef RP01 mutant upon expression of α-enolase (ENO1) and VDAC1. Data are presented as the mean ± SD of two independent experiment.

    Article Snippet: Passive Lysis Buffer (Promega), PVDF membrane (Millipore), Luminata forte Western HRP substrate (Millipore), anti alpha-tubulin monoclonal antibody (Invitrogen Life Technologies), DAPI (Invitrogen Life Technologies), Primary antibodies- anti-Cyclophilin A antibody (ab58144), anti-eIF5A antibody [EP527Y] (ab32407), anti-ENO1 antibody (ab85086), anti-OTUB1 antibody (ab98280), anti-RhoGDI antibody [1F2] (ab118159), and anti-VDAC1/Porin antibody—N-terminal (ab135585) were purchased from abcam, UK.

    Techniques: Western Blot, Mutagenesis, Expressing

    Predicted network map for Cyclophilin A, EIF5A-1, Rho GDI, VDAC1, OTUB1, and ENO1 with their top partners and association with HIV-1 Nef was created in STRING. Proteins in boxes were underexpressed proteins identified in our work. Rest of proteins were host proteins from the database. Protein labeled in Yellow is Nef, with known interaction with Cyclophilin A, RAC1, CDC42 and BCL2L1 (connected with red line) as given in HIV-1, Human Protein Interaction Database.

    Journal: PLoS ONE

    Article Title: Proteomic Profiling of SupT1 Cells Reveal Modulation of Host Proteins by HIV-1 Nef Variants

    doi: 10.1371/journal.pone.0122994

    Figure Lengend Snippet: Predicted network map for Cyclophilin A, EIF5A-1, Rho GDI, VDAC1, OTUB1, and ENO1 with their top partners and association with HIV-1 Nef was created in STRING. Proteins in boxes were underexpressed proteins identified in our work. Rest of proteins were host proteins from the database. Protein labeled in Yellow is Nef, with known interaction with Cyclophilin A, RAC1, CDC42 and BCL2L1 (connected with red line) as given in HIV-1, Human Protein Interaction Database.

    Article Snippet: Passive Lysis Buffer (Promega), PVDF membrane (Millipore), Luminata forte Western HRP substrate (Millipore), anti alpha-tubulin monoclonal antibody (Invitrogen Life Technologies), DAPI (Invitrogen Life Technologies), Primary antibodies- anti-Cyclophilin A antibody (ab58144), anti-eIF5A antibody [EP527Y] (ab32407), anti-ENO1 antibody (ab85086), anti-OTUB1 antibody (ab98280), anti-RhoGDI antibody [1F2] (ab118159), and anti-VDAC1/Porin antibody—N-terminal (ab135585) were purchased from abcam, UK.

    Techniques: Labeling