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polyglutamine lengths  (Addgene inc)


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    Addgene inc polyglutamine lengths
    Figure 2. Polythreonine forms detergent-insoluble aggregates (A) GFPs with a polyalanine or polythreonine tail were expressed under the GAL1 promoter in WT cells. Cell lysates were analyzed by SDS-PAGE, followed by anti- GFP immunoblotting (IB). Pgk1 served as a loading control. HMW, high-molecular-weight smear. (B) Fluorescence microscopy of WT cells expressing GFPs with the indicated C-terminal tails. White dashed lines indicate cell boundaries. BF, bright field. Scale bar, 5 mm. (C) FRAP analysis of GFP-T30 inclusions. Representative images of pre- and post-bleach are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-T20 was used as a control. (D) GFPs with C-terminal polythreonine tails were expressed under the GAL1 promoter in WT cells. Detergent-insoluble aggregates tended to stack within polyacrylamide gel pockets. The aggregates were better resolved using SDD-AGE. (E) Fluorescence microscopy of GFPs with polythreonine tails as described in (D). Representative inclusions are shown if detectable. Scale bar, 5 mm. The number of cells with GFP inclusions was counted and is represented in the graph as a percentage of GFP-expressing cells. Mean ± SEM from three biological replicates are shown, with more than 150 cells observed for each tail per biological replicate. (F) FLAG-tagged polythreonine peptides were expressed under the GAL1 promoter in WT cells. Fixed cells were spheroplasted and subjected to immunoflu- orescence microscopy using anti-FLAG antibody (a-FLAG). Scale bar, 5 mm. (G) WT cells expressing FLAG-tagged polythreonine peptides, as described in (F), were lysed and subjected to SDD-AGE and SDS-PAGE, followed by anti-FLAG immunoblotting. (H) Fluorescence microscopy of WT cells expressing GFPs with polythreonine, <t>polyglutamine,</t> and polyasparagine tails. Scale bar, 5 mm.
    Polyglutamine Lengths, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Threonine-rich carboxyl-terminal extension drives aggregation of stalled polypeptides."

    Article Title: Threonine-rich carboxyl-terminal extension drives aggregation of stalled polypeptides.

    Journal: Molecular cell

    doi: 10.1016/j.molcel.2024.10.011

    Figure 2. Polythreonine forms detergent-insoluble aggregates (A) GFPs with a polyalanine or polythreonine tail were expressed under the GAL1 promoter in WT cells. Cell lysates were analyzed by SDS-PAGE, followed by anti- GFP immunoblotting (IB). Pgk1 served as a loading control. HMW, high-molecular-weight smear. (B) Fluorescence microscopy of WT cells expressing GFPs with the indicated C-terminal tails. White dashed lines indicate cell boundaries. BF, bright field. Scale bar, 5 mm. (C) FRAP analysis of GFP-T30 inclusions. Representative images of pre- and post-bleach are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-T20 was used as a control. (D) GFPs with C-terminal polythreonine tails were expressed under the GAL1 promoter in WT cells. Detergent-insoluble aggregates tended to stack within polyacrylamide gel pockets. The aggregates were better resolved using SDD-AGE. (E) Fluorescence microscopy of GFPs with polythreonine tails as described in (D). Representative inclusions are shown if detectable. Scale bar, 5 mm. The number of cells with GFP inclusions was counted and is represented in the graph as a percentage of GFP-expressing cells. Mean ± SEM from three biological replicates are shown, with more than 150 cells observed for each tail per biological replicate. (F) FLAG-tagged polythreonine peptides were expressed under the GAL1 promoter in WT cells. Fixed cells were spheroplasted and subjected to immunoflu- orescence microscopy using anti-FLAG antibody (a-FLAG). Scale bar, 5 mm. (G) WT cells expressing FLAG-tagged polythreonine peptides, as described in (F), were lysed and subjected to SDD-AGE and SDS-PAGE, followed by anti-FLAG immunoblotting. (H) Fluorescence microscopy of WT cells expressing GFPs with polythreonine, polyglutamine, and polyasparagine tails. Scale bar, 5 mm.
    Figure Legend Snippet: Figure 2. Polythreonine forms detergent-insoluble aggregates (A) GFPs with a polyalanine or polythreonine tail were expressed under the GAL1 promoter in WT cells. Cell lysates were analyzed by SDS-PAGE, followed by anti- GFP immunoblotting (IB). Pgk1 served as a loading control. HMW, high-molecular-weight smear. (B) Fluorescence microscopy of WT cells expressing GFPs with the indicated C-terminal tails. White dashed lines indicate cell boundaries. BF, bright field. Scale bar, 5 mm. (C) FRAP analysis of GFP-T30 inclusions. Representative images of pre- and post-bleach are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-T20 was used as a control. (D) GFPs with C-terminal polythreonine tails were expressed under the GAL1 promoter in WT cells. Detergent-insoluble aggregates tended to stack within polyacrylamide gel pockets. The aggregates were better resolved using SDD-AGE. (E) Fluorescence microscopy of GFPs with polythreonine tails as described in (D). Representative inclusions are shown if detectable. Scale bar, 5 mm. The number of cells with GFP inclusions was counted and is represented in the graph as a percentage of GFP-expressing cells. Mean ± SEM from three biological replicates are shown, with more than 150 cells observed for each tail per biological replicate. (F) FLAG-tagged polythreonine peptides were expressed under the GAL1 promoter in WT cells. Fixed cells were spheroplasted and subjected to immunoflu- orescence microscopy using anti-FLAG antibody (a-FLAG). Scale bar, 5 mm. (G) WT cells expressing FLAG-tagged polythreonine peptides, as described in (F), were lysed and subjected to SDD-AGE and SDS-PAGE, followed by anti-FLAG immunoblotting. (H) Fluorescence microscopy of WT cells expressing GFPs with polythreonine, polyglutamine, and polyasparagine tails. Scale bar, 5 mm.

    Techniques Used: SDS Page, Western Blot, Control, High Molecular Weight, Fluorescence, Microscopy, Expressing

    Figure 5. Polythreonine aggregation in human cells (A) HeLa cells were transfected to express GFPs containing either polyalanine, polythreonine, polyglutamine, or polyserine tails. Nuclei were counterstained with Hoechst and represented in gray in merged images. Scale bar, 15 mm. (B) FRAP analyses of GFP, GFP-T30, and Htt72Q-GFP (huntingtin exon 1 containing a 72-residue-long polyglutamine tract) in HeLa cells. Representative images of GFP-T30 and Htt72Q-GFP inclusions are shown, with photobleached areas circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. (C) Lysates of HeLa cells expressing GFPs, with or without the indicated polyamino acid tails, were analyzed by anti-GFP immunoblotting (IB). b-actin served as a loading control. (D) Fluorescence microscopy of GFP or GFP-S30, co-expressed with BFP or BFP-T30 in HeLa cells. Scale bar, 15 mm. (E) FRAP analyses of GFP-S30 co-localized with BFP-T30 inclusions in HeLa cells. Representative images of inclusions are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-S30 co-expressed with BFP was used as a control. (F) Lysates of HeLa cells expressing GFP-S30 (or GFP) and BFP-T30 (or BFP) were analyzed by SDD-AGE and SDS-PAGE.
    Figure Legend Snippet: Figure 5. Polythreonine aggregation in human cells (A) HeLa cells were transfected to express GFPs containing either polyalanine, polythreonine, polyglutamine, or polyserine tails. Nuclei were counterstained with Hoechst and represented in gray in merged images. Scale bar, 15 mm. (B) FRAP analyses of GFP, GFP-T30, and Htt72Q-GFP (huntingtin exon 1 containing a 72-residue-long polyglutamine tract) in HeLa cells. Representative images of GFP-T30 and Htt72Q-GFP inclusions are shown, with photobleached areas circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. (C) Lysates of HeLa cells expressing GFPs, with or without the indicated polyamino acid tails, were analyzed by anti-GFP immunoblotting (IB). b-actin served as a loading control. (D) Fluorescence microscopy of GFP or GFP-S30, co-expressed with BFP or BFP-T30 in HeLa cells. Scale bar, 15 mm. (E) FRAP analyses of GFP-S30 co-localized with BFP-T30 inclusions in HeLa cells. Representative images of inclusions are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-S30 co-expressed with BFP was used as a control. (F) Lysates of HeLa cells expressing GFP-S30 (or GFP) and BFP-T30 (or BFP) were analyzed by SDD-AGE and SDS-PAGE.

    Techniques Used: Transfection, Residue, Expressing, Western Blot, Control, Fluorescence, Microscopy, SDS Page



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    Figure 2. Polythreonine forms detergent-insoluble aggregates (A) GFPs with a polyalanine or polythreonine tail were expressed under the GAL1 promoter in WT cells. Cell lysates were analyzed by SDS-PAGE, followed by anti- GFP immunoblotting (IB). Pgk1 served as a loading control. HMW, high-molecular-weight smear. (B) Fluorescence microscopy of WT cells expressing GFPs with the indicated C-terminal tails. White dashed lines indicate cell boundaries. BF, bright field. Scale bar, 5 mm. (C) FRAP analysis of GFP-T30 inclusions. Representative images of pre- and post-bleach are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-T20 was used as a control. (D) GFPs with C-terminal polythreonine tails were expressed under the GAL1 promoter in WT cells. Detergent-insoluble aggregates tended to stack within polyacrylamide gel pockets. The aggregates were better resolved using SDD-AGE. (E) Fluorescence microscopy of GFPs with polythreonine tails as described in (D). Representative inclusions are shown if detectable. Scale bar, 5 mm. The number of cells with GFP inclusions was counted and is represented in the graph as a percentage of GFP-expressing cells. Mean ± SEM from three biological replicates are shown, with more than 150 cells observed for each tail per biological replicate. (F) FLAG-tagged polythreonine peptides were expressed under the GAL1 promoter in WT cells. Fixed cells were spheroplasted and subjected to immunoflu- orescence microscopy using anti-FLAG antibody (a-FLAG). Scale bar, 5 mm. (G) WT cells expressing FLAG-tagged polythreonine peptides, as described in (F), were lysed and subjected to SDD-AGE and SDS-PAGE, followed by anti-FLAG immunoblotting. (H) Fluorescence microscopy of WT cells expressing GFPs with polythreonine, <t>polyglutamine,</t> and polyasparagine tails. Scale bar, 5 mm.
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    Image Search Results


    Figure 2. Polythreonine forms detergent-insoluble aggregates (A) GFPs with a polyalanine or polythreonine tail were expressed under the GAL1 promoter in WT cells. Cell lysates were analyzed by SDS-PAGE, followed by anti- GFP immunoblotting (IB). Pgk1 served as a loading control. HMW, high-molecular-weight smear. (B) Fluorescence microscopy of WT cells expressing GFPs with the indicated C-terminal tails. White dashed lines indicate cell boundaries. BF, bright field. Scale bar, 5 mm. (C) FRAP analysis of GFP-T30 inclusions. Representative images of pre- and post-bleach are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-T20 was used as a control. (D) GFPs with C-terminal polythreonine tails were expressed under the GAL1 promoter in WT cells. Detergent-insoluble aggregates tended to stack within polyacrylamide gel pockets. The aggregates were better resolved using SDD-AGE. (E) Fluorescence microscopy of GFPs with polythreonine tails as described in (D). Representative inclusions are shown if detectable. Scale bar, 5 mm. The number of cells with GFP inclusions was counted and is represented in the graph as a percentage of GFP-expressing cells. Mean ± SEM from three biological replicates are shown, with more than 150 cells observed for each tail per biological replicate. (F) FLAG-tagged polythreonine peptides were expressed under the GAL1 promoter in WT cells. Fixed cells were spheroplasted and subjected to immunoflu- orescence microscopy using anti-FLAG antibody (a-FLAG). Scale bar, 5 mm. (G) WT cells expressing FLAG-tagged polythreonine peptides, as described in (F), were lysed and subjected to SDD-AGE and SDS-PAGE, followed by anti-FLAG immunoblotting. (H) Fluorescence microscopy of WT cells expressing GFPs with polythreonine, polyglutamine, and polyasparagine tails. Scale bar, 5 mm.

    Journal: Molecular cell

    Article Title: Threonine-rich carboxyl-terminal extension drives aggregation of stalled polypeptides.

    doi: 10.1016/j.molcel.2024.10.011

    Figure Lengend Snippet: Figure 2. Polythreonine forms detergent-insoluble aggregates (A) GFPs with a polyalanine or polythreonine tail were expressed under the GAL1 promoter in WT cells. Cell lysates were analyzed by SDS-PAGE, followed by anti- GFP immunoblotting (IB). Pgk1 served as a loading control. HMW, high-molecular-weight smear. (B) Fluorescence microscopy of WT cells expressing GFPs with the indicated C-terminal tails. White dashed lines indicate cell boundaries. BF, bright field. Scale bar, 5 mm. (C) FRAP analysis of GFP-T30 inclusions. Representative images of pre- and post-bleach are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-T20 was used as a control. (D) GFPs with C-terminal polythreonine tails were expressed under the GAL1 promoter in WT cells. Detergent-insoluble aggregates tended to stack within polyacrylamide gel pockets. The aggregates were better resolved using SDD-AGE. (E) Fluorescence microscopy of GFPs with polythreonine tails as described in (D). Representative inclusions are shown if detectable. Scale bar, 5 mm. The number of cells with GFP inclusions was counted and is represented in the graph as a percentage of GFP-expressing cells. Mean ± SEM from three biological replicates are shown, with more than 150 cells observed for each tail per biological replicate. (F) FLAG-tagged polythreonine peptides were expressed under the GAL1 promoter in WT cells. Fixed cells were spheroplasted and subjected to immunoflu- orescence microscopy using anti-FLAG antibody (a-FLAG). Scale bar, 5 mm. (G) WT cells expressing FLAG-tagged polythreonine peptides, as described in (F), were lysed and subjected to SDD-AGE and SDS-PAGE, followed by anti-FLAG immunoblotting. (H) Fluorescence microscopy of WT cells expressing GFPs with polythreonine, polyglutamine, and polyasparagine tails. Scale bar, 5 mm.

    Article Snippet: Vectors for C-terminal GFP-tagged Huntingtin proteins of various polyglutamine lengths (Htt25Q, Htt46Q, Htt72Q and Htt103Q) were obtained from Addgene (#1185, #15581, #15582 and #1186, respectively) and used directly without modification in yeast.

    Techniques: SDS Page, Western Blot, Control, High Molecular Weight, Fluorescence, Microscopy, Expressing

    Figure 5. Polythreonine aggregation in human cells (A) HeLa cells were transfected to express GFPs containing either polyalanine, polythreonine, polyglutamine, or polyserine tails. Nuclei were counterstained with Hoechst and represented in gray in merged images. Scale bar, 15 mm. (B) FRAP analyses of GFP, GFP-T30, and Htt72Q-GFP (huntingtin exon 1 containing a 72-residue-long polyglutamine tract) in HeLa cells. Representative images of GFP-T30 and Htt72Q-GFP inclusions are shown, with photobleached areas circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. (C) Lysates of HeLa cells expressing GFPs, with or without the indicated polyamino acid tails, were analyzed by anti-GFP immunoblotting (IB). b-actin served as a loading control. (D) Fluorescence microscopy of GFP or GFP-S30, co-expressed with BFP or BFP-T30 in HeLa cells. Scale bar, 15 mm. (E) FRAP analyses of GFP-S30 co-localized with BFP-T30 inclusions in HeLa cells. Representative images of inclusions are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-S30 co-expressed with BFP was used as a control. (F) Lysates of HeLa cells expressing GFP-S30 (or GFP) and BFP-T30 (or BFP) were analyzed by SDD-AGE and SDS-PAGE.

    Journal: Molecular cell

    Article Title: Threonine-rich carboxyl-terminal extension drives aggregation of stalled polypeptides.

    doi: 10.1016/j.molcel.2024.10.011

    Figure Lengend Snippet: Figure 5. Polythreonine aggregation in human cells (A) HeLa cells were transfected to express GFPs containing either polyalanine, polythreonine, polyglutamine, or polyserine tails. Nuclei were counterstained with Hoechst and represented in gray in merged images. Scale bar, 15 mm. (B) FRAP analyses of GFP, GFP-T30, and Htt72Q-GFP (huntingtin exon 1 containing a 72-residue-long polyglutamine tract) in HeLa cells. Representative images of GFP-T30 and Htt72Q-GFP inclusions are shown, with photobleached areas circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. (C) Lysates of HeLa cells expressing GFPs, with or without the indicated polyamino acid tails, were analyzed by anti-GFP immunoblotting (IB). b-actin served as a loading control. (D) Fluorescence microscopy of GFP or GFP-S30, co-expressed with BFP or BFP-T30 in HeLa cells. Scale bar, 15 mm. (E) FRAP analyses of GFP-S30 co-localized with BFP-T30 inclusions in HeLa cells. Representative images of inclusions are shown, with the photobleached area circled. Scale bar, 1 mm. Graph curves represent mean measurements of 10 inclusions, with error bars indicating SEM. Diffuse GFP-S30 co-expressed with BFP was used as a control. (F) Lysates of HeLa cells expressing GFP-S30 (or GFP) and BFP-T30 (or BFP) were analyzed by SDD-AGE and SDS-PAGE.

    Article Snippet: Vectors for C-terminal GFP-tagged Huntingtin proteins of various polyglutamine lengths (Htt25Q, Htt46Q, Htt72Q and Htt103Q) were obtained from Addgene (#1185, #15581, #15582 and #1186, respectively) and used directly without modification in yeast.

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