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pmcherry c1 eps8 plasmid  (Addgene inc)


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    Structured Review

    Addgene inc pmcherry c1 eps8 plasmid
    MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) <t>mCherry-EPS8</t> or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.
    Pmcherry C1 Eps8 Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmcherry c1 eps8 plasmid/product/Addgene inc
    Average 90 stars, based on 6 article reviews
    pmcherry c1 eps8 plasmid - by Bioz Stars, 2026-02
    90/100 stars

    Images

    1) Product Images from "A Myosin Nanomotor Essential for Stereocilia Maintenance Expands the Etiology of Hereditary Hearing Loss DFNB3"

    Article Title: A Myosin Nanomotor Essential for Stereocilia Maintenance Expands the Etiology of Hereditary Hearing Loss DFNB3

    Journal: bioRxiv

    doi: 10.1101/2025.02.19.639121

    MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) mCherry-EPS8 or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.
    Figure Legend Snippet: MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) mCherry-EPS8 or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.

    Techniques Used: Transfection, Positive Control, Negative Control, Microscopy, Fluorescence

    MYO15A-3 is not required for the elongation complex trafficking in hair cells at P4, but traffics the elongation complex at P21. (A, C) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 4 (P4) immunolabeled for the EPS8/ WHRN (green). The hair cells were counterstained with phalloidin (magenta) to visualize the actin-rich stereocilia bundles. (B, D) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Each dot represents the fluorescence intensity of a single hair cell. This was measured by averaging the absolute fluorescence intensity at the tips of individual row 1 stereocilia. Data are mean ± SD for n=3 mice per genotype (total ∼33-38 hair cells per condition/genotype). The absolute fluorescence for EPS8/WHRN was not significantly different between genotypes, as determined via the Mann-Whitney U-test to measure EPS8 level and an unpaired t-test for WHRN. (E, G) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 21 (P21) immunolabeled for the EPS8/ WHRN (green). (F, H) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Data are mean ± SD for n=3 mice per genotype (total ∼28-35 hair cells per condition/genotype) with statistical significance of **** P<0.0001 via Mann-Whitney U-test to measure WHRN and an unpaired t-test for EPS8. Scale bars: 5 µm.
    Figure Legend Snippet: MYO15A-3 is not required for the elongation complex trafficking in hair cells at P4, but traffics the elongation complex at P21. (A, C) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 4 (P4) immunolabeled for the EPS8/ WHRN (green). The hair cells were counterstained with phalloidin (magenta) to visualize the actin-rich stereocilia bundles. (B, D) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Each dot represents the fluorescence intensity of a single hair cell. This was measured by averaging the absolute fluorescence intensity at the tips of individual row 1 stereocilia. Data are mean ± SD for n=3 mice per genotype (total ∼33-38 hair cells per condition/genotype). The absolute fluorescence for EPS8/WHRN was not significantly different between genotypes, as determined via the Mann-Whitney U-test to measure EPS8 level and an unpaired t-test for WHRN. (E, G) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 21 (P21) immunolabeled for the EPS8/ WHRN (green). (F, H) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Data are mean ± SD for n=3 mice per genotype (total ∼28-35 hair cells per condition/genotype) with statistical significance of **** P<0.0001 via Mann-Whitney U-test to measure WHRN and an unpaired t-test for EPS8. Scale bars: 5 µm.

    Techniques Used: Immunolabeling, Fluorescence, MANN-WHITNEY



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    MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) <t>mCherry-EPS8</t> or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.
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    MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) <t>mCherry-EPS8</t> or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.
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    MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) <t>mCherry-EPS8</t> or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.
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    Image Search Results


    MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) mCherry-EPS8 or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.

    Journal: bioRxiv

    Article Title: A Myosin Nanomotor Essential for Stereocilia Maintenance Expands the Etiology of Hereditary Hearing Loss DFNB3

    doi: 10.1101/2025.02.19.639121

    Figure Lengend Snippet: MYO15A-3 traffics the elongation complex protein in filopodia. HeLa cells were transfected with plasmids encoding for protein fusion proteins, either EGFP-MYO15A-2 (positive control), EGFP-MYO15A-3 or EGFP-MYO10 (negative control), and co-transfected with either (A-C) mCherry-EPS8 or (H-J) mCherry-WHRN to detect endogenous protein. Cells were imaged using a spinning disk confocal microscope to investigate the localization and interaction of these proteins. The intensity of EGFP-Myosin and mCherry-EPS8/WHRN was measured blinded at multiple filopodia tips (∼300 filopodia from n=3 independent transfections). (D-F, K-M) The molecular interaction between the myosin and the EC proteins was determined using Pearson’s r coefficient, calculated along the filopodia shaft to measure the fluorescence correlation between the bait and prey. (G, N) Each data point represents the average interaction index from a single experiment (3 independent determinations) with a total of 30 cells per condition. The data are presented as mean ± SD with statistical significance determined using One-way ANOVA. *** P<0.001, **** P<0.0001. Scale bars: 5 µm.

    Article Snippet: The pmCherry-C1-EPS8 plasmid was a gift from Christien Merrifield (Addgene plasmid #29779).

    Techniques: Transfection, Positive Control, Negative Control, Microscopy, Fluorescence

    MYO15A-3 is not required for the elongation complex trafficking in hair cells at P4, but traffics the elongation complex at P21. (A, C) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 4 (P4) immunolabeled for the EPS8/ WHRN (green). The hair cells were counterstained with phalloidin (magenta) to visualize the actin-rich stereocilia bundles. (B, D) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Each dot represents the fluorescence intensity of a single hair cell. This was measured by averaging the absolute fluorescence intensity at the tips of individual row 1 stereocilia. Data are mean ± SD for n=3 mice per genotype (total ∼33-38 hair cells per condition/genotype). The absolute fluorescence for EPS8/WHRN was not significantly different between genotypes, as determined via the Mann-Whitney U-test to measure EPS8 level and an unpaired t-test for WHRN. (E, G) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 21 (P21) immunolabeled for the EPS8/ WHRN (green). (F, H) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Data are mean ± SD for n=3 mice per genotype (total ∼28-35 hair cells per condition/genotype) with statistical significance of **** P<0.0001 via Mann-Whitney U-test to measure WHRN and an unpaired t-test for EPS8. Scale bars: 5 µm.

    Journal: bioRxiv

    Article Title: A Myosin Nanomotor Essential for Stereocilia Maintenance Expands the Etiology of Hereditary Hearing Loss DFNB3

    doi: 10.1101/2025.02.19.639121

    Figure Lengend Snippet: MYO15A-3 is not required for the elongation complex trafficking in hair cells at P4, but traffics the elongation complex at P21. (A, C) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 4 (P4) immunolabeled for the EPS8/ WHRN (green). The hair cells were counterstained with phalloidin (magenta) to visualize the actin-rich stereocilia bundles. (B, D) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Each dot represents the fluorescence intensity of a single hair cell. This was measured by averaging the absolute fluorescence intensity at the tips of individual row 1 stereocilia. Data are mean ± SD for n=3 mice per genotype (total ∼33-38 hair cells per condition/genotype). The absolute fluorescence for EPS8/WHRN was not significantly different between genotypes, as determined via the Mann-Whitney U-test to measure EPS8 level and an unpaired t-test for WHRN. (E, G) Confocal images of hair cells from cochleae of Myo15a (+/ Δ 3) and Myo15a ( Δ 3/ Δ 3) mice at postnatal day 21 (P21) immunolabeled for the EPS8/ WHRN (green). (F, H) Quantification of EPS8 and WHRN relative intensity at the stereocilia tips. Data are mean ± SD for n=3 mice per genotype (total ∼28-35 hair cells per condition/genotype) with statistical significance of **** P<0.0001 via Mann-Whitney U-test to measure WHRN and an unpaired t-test for EPS8. Scale bars: 5 µm.

    Article Snippet: The pmCherry-C1-EPS8 plasmid was a gift from Christien Merrifield (Addgene plasmid #29779).

    Techniques: Immunolabeling, Fluorescence, MANN-WHITNEY