anti mapk7 antibody  (Boster Bio)


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    Boster Bio anti mapk7 antibody
    IR-induced p90RSK activation lead to <t>ERK5</t> S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).
    Anti Mapk7 Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mapk7 antibody/product/Boster Bio
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti mapk7 antibody - by Bioz Stars, 2022-12
    90/100 stars

    Images

    1) Product Images from "Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation"

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    Journal: Frontiers in Cardiovascular Medicine

    doi: 10.3389/fcvm.2018.00023

    IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).
    Figure Legend Snippet: IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).

    Techniques Used: Activation Assay, Activity Assay, Western Blot, Microelectrode Array

    ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)
    Figure Legend Snippet: ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)

    Techniques Used: Transfection, Luciferase, Plasmid Preparation, Activity Assay, Expressing, Mutagenesis, Activation Assay, Staining, Flow Cytometry, Cytometry

    ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.
    Figure Legend Snippet: ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Mutagenesis, Flow Cytometry, Cytometry

    2) Product Images from "Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation"

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    Journal: Frontiers in Cardiovascular Medicine

    doi: 10.3389/fcvm.2018.00023

    IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).
    Figure Legend Snippet: IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).

    Techniques Used: Activation Assay, Activity Assay, Western Blot, Microelectrode Array

    ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)
    Figure Legend Snippet: ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)

    Techniques Used: Transfection, Luciferase, Plasmid Preparation, Activity Assay, Expressing, Mutagenesis, Activation Assay, Staining, Flow Cytometry, Cytometry

    ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.
    Figure Legend Snippet: ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Mutagenesis, Flow Cytometry, Cytometry

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    Boster Bio anti mapk7 antibody
    IR-induced p90RSK activation lead to <t>ERK5</t> S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).
    Anti Mapk7 Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mapk7 antibody/product/Boster Bio
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mapk7 antibody - by Bioz Stars, 2022-12
    90/100 stars
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    IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    doi: 10.3389/fcvm.2018.00023

    Figure Lengend Snippet: IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).

    Article Snippet: Antibodies and Reagents Antibodies were purchased from the following vendors: anti-p90RSK1 (C-21, #SC-231) and anti-VCAM-1 (SC-8304) from Santa Cruz Biotechnology (Dallas, TX); anti-phospho-p90RSK (Ser380, #9341), anti-phospho-ERK5 (Thr218/Tyr220, #3371L), and anti-ERK5 (#3372) from Cell Signaling (Danvers, MA); anti-tubulin (T-5168) from Sigma (St. Louis, MO); anti-VE-cadherin (#555289) from BD Transduction Laboratory (San Diego, CA); and anti-phopho-specific ERK5 S496 antibody (#A02812) from Boster Bio (Pleasanton, CA).

    Techniques: Activation Assay, Activity Assay, Western Blot, Microelectrode Array

    ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    doi: 10.3389/fcvm.2018.00023

    Figure Lengend Snippet: ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)

    Article Snippet: Antibodies and Reagents Antibodies were purchased from the following vendors: anti-p90RSK1 (C-21, #SC-231) and anti-VCAM-1 (SC-8304) from Santa Cruz Biotechnology (Dallas, TX); anti-phospho-p90RSK (Ser380, #9341), anti-phospho-ERK5 (Thr218/Tyr220, #3371L), and anti-ERK5 (#3372) from Cell Signaling (Danvers, MA); anti-tubulin (T-5168) from Sigma (St. Louis, MO); anti-VE-cadherin (#555289) from BD Transduction Laboratory (San Diego, CA); and anti-phopho-specific ERK5 S496 antibody (#A02812) from Boster Bio (Pleasanton, CA).

    Techniques: Transfection, Luciferase, Plasmid Preparation, Activity Assay, Expressing, Mutagenesis, Activation Assay, Staining, Flow Cytometry, Cytometry

    ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    doi: 10.3389/fcvm.2018.00023

    Figure Lengend Snippet: ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.

    Article Snippet: Antibodies and Reagents Antibodies were purchased from the following vendors: anti-p90RSK1 (C-21, #SC-231) and anti-VCAM-1 (SC-8304) from Santa Cruz Biotechnology (Dallas, TX); anti-phospho-p90RSK (Ser380, #9341), anti-phospho-ERK5 (Thr218/Tyr220, #3371L), and anti-ERK5 (#3372) from Cell Signaling (Danvers, MA); anti-tubulin (T-5168) from Sigma (St. Louis, MO); anti-VE-cadherin (#555289) from BD Transduction Laboratory (San Diego, CA); and anti-phopho-specific ERK5 S496 antibody (#A02812) from Boster Bio (Pleasanton, CA).

    Techniques: Transfection, Expressing, Plasmid Preparation, Mutagenesis, Flow Cytometry, Cytometry

    IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    doi: 10.3389/fcvm.2018.00023

    Figure Lengend Snippet: IR-induced p90RSK activation lead to ERK5 S496 phosphorylation, but not ERK5 TEY motif phosphorylation. ( A ) HUVECs were treated with IR by the indicated doses, and 30 min after IR, ECs were collected and p90RSK activity was detected by Western blotting with anti–p-p90RSK and anti-p90RSK antibodies. Representative images from three independent experiments are shown. ( B ) HAECs were pre-treated with FMK-MEA (5 µM) for 30 min and treated by IR (2 Gy) for the indicated times. ECs were collected, and total p90RSK, p90RSK phosphorylation, ERK5 S496 phosphorylation, ERK5 TEY motif phosphorylation, and total ERK5 were detected by Western blotting with anti-p90RSK, anti-phospho-p90RSK (S380), anti-phospho-ERK5 (S496), anti-phospho-ERK5 (TEY motif), and anti-ERK5. Representative images from three independent experiments are shown. ( C ) Quantification of IR-induced p90RSK activation (S380 phosphorylation; top), ERK5 S496 phosphorylation (middle), and ERK5 activation (TEY motif phosphorylation; bottom) is shown after normalization by total protein levels. Data represent mean ± SEM ( n = 3).

    Article Snippet: Antibodies were purchased from the following vendors: anti-p90RSK1 (C-21, #SC-231) and anti-VCAM-1 (SC-8304) from Santa Cruz Biotechnology (Dallas, TX); anti-phospho-p90RSK (Ser380, #9341), anti-phospho-ERK5 (Thr218/Tyr220, #3371L), and anti-ERK5 (#3372) from Cell Signaling (Danvers, MA); anti-tubulin (T-5168) from Sigma (St. Louis, MO); anti-VE-cadherin (#555289) from BD Transduction Laboratory (San Diego, CA); and anti-phopho-specific ERK5 S496 antibody (#A02812) from Boster Bio (Pleasanton, CA).

    Techniques: Activation Assay, Activity Assay, Western Blot, Microelectrode Array

    ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    doi: 10.3389/fcvm.2018.00023

    Figure Lengend Snippet: ERK5 S496 phosphorylation is critical for IR-induced EC inflammation. HUVECs were co-transfected with the NF-κB-Luc luciferase reporter vector and the pRL-CMV vector (renilla luciferase activity, internal control) together with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy) and harvested after 24 h. ( A ) NF-κB activation was quantified by measuring relative luciferase activity using a dual-luciferase reporter system. Shown are relative luciferase activity, presented as firefly luciferase/renilla luciferase activity ratio. ( n = 6) ( B ) VCAM1 expression was determined by measuring percentage of positive VCAM1-staining cells using the BD Accuri C6 Flow Cytometer system using the FL1 533/30 nm filter. ( n = 5)

    Article Snippet: Antibodies were purchased from the following vendors: anti-p90RSK1 (C-21, #SC-231) and anti-VCAM-1 (SC-8304) from Santa Cruz Biotechnology (Dallas, TX); anti-phospho-p90RSK (Ser380, #9341), anti-phospho-ERK5 (Thr218/Tyr220, #3371L), and anti-ERK5 (#3372) from Cell Signaling (Danvers, MA); anti-tubulin (T-5168) from Sigma (St. Louis, MO); anti-VE-cadherin (#555289) from BD Transduction Laboratory (San Diego, CA); and anti-phopho-specific ERK5 S496 antibody (#A02812) from Boster Bio (Pleasanton, CA).

    Techniques: Transfection, Luciferase, Plasmid Preparation, Activity Assay, Expressing, Mutagenesis, Activation Assay, Staining, Flow Cytometry, Cytometry

    ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Ionizing Radiation Induces Endothelial Inflammation and Apoptosis via p90RSK-Mediated ERK5 S496 Phosphorylation

    doi: 10.3389/fcvm.2018.00023

    Figure Lengend Snippet: ERK5 S496 phosphorylation is critical for IR-induced apoptosis. HUVECs were transfected with the expression vector encoding the flag-tagged ERK5 wild type (WT) or the S496A mutant. Transfected cells were then exposed to gamma radiation (2 Gy), and harvested after 60 h. Cell apoptosis was determined by measuring percentage of Annexin V- positive cells (Abcam’s Annexin V-FITC Apoptosis Detection Kit) using the BD Accuri C6 Flow Cytometer system with the FL1 533/30 nm filter. Shown are mean ± SEM, n = 3–6.

    Article Snippet: Antibodies were purchased from the following vendors: anti-p90RSK1 (C-21, #SC-231) and anti-VCAM-1 (SC-8304) from Santa Cruz Biotechnology (Dallas, TX); anti-phospho-p90RSK (Ser380, #9341), anti-phospho-ERK5 (Thr218/Tyr220, #3371L), and anti-ERK5 (#3372) from Cell Signaling (Danvers, MA); anti-tubulin (T-5168) from Sigma (St. Louis, MO); anti-VE-cadherin (#555289) from BD Transduction Laboratory (San Diego, CA); and anti-phopho-specific ERK5 S496 antibody (#A02812) from Boster Bio (Pleasanton, CA).

    Techniques: Transfection, Expressing, Plasmid Preparation, Mutagenesis, Flow Cytometry, Cytometry