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e64d  (Tocris)


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    Tocris e64d
    Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and <t>E64d</t> targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.
    E64d, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e64d/product/Tocris
    Average 94 stars, based on 45 article reviews
    e64d - by Bioz Stars, 2026-04
    94/100 stars

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    1) Product Images from "The impact of clade B lineage 5 MERS coronaviruses spike mutations from 2015 to 2023 on virus entry and replication competence"

    Article Title: The impact of clade B lineage 5 MERS coronaviruses spike mutations from 2015 to 2023 on virus entry and replication competence

    Journal: PLOS Pathogens

    doi: 10.1371/journal.ppat.1013336

    Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and E64d targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.
    Figure Legend Snippet: Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and E64d targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.

    Techniques Used: Blocking Assay, Virus, Comparison, Inhibition, Quantitative RT-PCR, RNA Expression, Flow Cytometry, Expressing, Infection



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    Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and <t>E64d</t> targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.
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    Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and <t>E64d</t> targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.
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    Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and E64d targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.

    Journal: PLOS Pathogens

    Article Title: The impact of clade B lineage 5 MERS coronaviruses spike mutations from 2015 to 2023 on virus entry and replication competence

    doi: 10.1371/journal.ppat.1013336

    Figure Lengend Snippet: Treatment of cells with camostat targeting the cell surface TMPRSS2 pathway and E64d targeting the endocytic cathepsin L pathway was used to block infectious virus entry via each pathway. Experiments with infectious virus (A) or pseudovirus (B) is shown for the different spike variants in VeroE6/TMPRSS2 cells. (A) Relative viral load was measured the RNA copies of MERS-CoV using the UpE assay in the cell supernatant at 48 hpi. (B) Relative cell entry was measured by RLU. (A-B) 2015/GD01 was the reference for comparison. Figures show the representative data from 2 independent experiments each with two biological replicates for each experiment each drug dose. An inhibition curve was fitted using the Hill equation with inhibition in Graphpad Prism. (C) RT-qPCR to measure RNA expression of DPP4, TMPRSS2 and CTSL gene among difference cell lines. Expressions were normalized by GAPDH gene for each cell line and relative fold changes were compared to 293T cells as reference. Flow cytometry analysis of surface expression of DPP4 and TMPRSS2 among different cell lines. Background levels were determined by isotype controls as indicated in grey. (D) Caco2 cells representing the TMPRSS2 entry pathway model and Huh7 cells representing the endosomal entry pathway model were pretreated with Camostat (25 μM), E64d (10 μM) or in both. Pseudoviruses were infected on drug treated cells in quadruplicate wells. Data shown are a representative of two independent repeats.

    Article Snippet: 3193) and E64d (Tocris, Cat. No. 4545) were serially diluted with 2% FBS DMEM medium and incubated on to VeroE6/TMPRSS2 cells for 2 hours prior infection.

    Techniques: Blocking Assay, Virus, Comparison, Inhibition, Quantitative RT-PCR, RNA Expression, Flow Cytometry, Expressing, Infection