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gfp rab5 dn plasmid (Addgene inc)

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Addgene inc gfp rab5 dn plasmid

CPAP depletion disrupts Rab5-to-Rab7 conversion (A) HeLa cells expressing control or CPAP shRNA were treated with untagged EGF for 60 min, stained for Rab5 and Rab7, and imaged by Airyscan super-resolution microscopy. Left: maximum intensity-projection images of Z stacks; middle: single Z-plane of images; right: co-localization (yellow) was quantified by determining the percentages of Rab5-positive (green) puncta containing Rab7 (red) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. (B) HeLa cells expressing control or CPAP shRNA were transfected <t>with</t> <t>GFP-Rab5</t> and mCherry-Rab7 constructs, treated with untagged EGF for 60 min, and imaged by Airyscan super-resolution microscopy. Left: maximum intensity-projection images of Z stacks; middle: single Z-plane of images; right: co-localization (yellow) was quantified by determining the percentages of GFP-Rab5-positive (green) puncta containing mCherry-Rab7 (red) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. The object-based co-localization macro tool FIJI was employed. (C) HeLa cells stably expressing control or CPAP-shRNA were transfected with control vector (GFP), GFP-Rab7 (WT; wild-type) or GFP-Rab7 (DN; dominant negative) vector constructs, treated with AF555-EGF ligand for 60 min, and stained for CD63 to mark MVBs/late endosomes, and imaged by confocal microscopy to determine AF555-EGF and CD63 co-localization. Left top row: GFP expression in control and Rab7 construct-expressing cells; left bottom rows: representative single Z-plane of images showing ligand-bound EGFR-CD63 co-localization; right: co-localization (yellow) was quantified by determining the percentages of EGF-positive (red) puncta containing CD63 (green) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. Zoomed images correspond to the dashed inset boxes of the indicated images. Scale bars: 10 μm. p values: ∗∗∗∗<0.0001 by unpaired nonparametric Mann-Whitney test.

Gfp Rab5 Dn Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/gfp+rab5+dn+plasmid/pmc12861012-40-0-4?v=Addgene+inc
Average 92 stars, based on 15 article reviews

gfp rab5 dn plasmid - by Bioz Stars, 2026-07

92/100 stars

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1) Product Images from "Centrosomal P4.1-associated protein is a novel regulator of ESCRT pathway function during endosome maturation"

Article Title: Centrosomal P4.1-associated protein is a novel regulator of ESCRT pathway function during endosome maturation

Journal: iScience

doi: 10.1016/j.isci.2026.114659

Figure Legend Snippet: CPAP depletion disrupts Rab5-to-Rab7 conversion (A) HeLa cells expressing control or CPAP shRNA were treated with untagged EGF for 60 min, stained for Rab5 and Rab7, and imaged by Airyscan super-resolution microscopy. Left: maximum intensity-projection images of Z stacks; middle: single Z-plane of images; right: co-localization (yellow) was quantified by determining the percentages of Rab5-positive (green) puncta containing Rab7 (red) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. (B) HeLa cells expressing control or CPAP shRNA were transfected with GFP-Rab5 and mCherry-Rab7 constructs, treated with untagged EGF for 60 min, and imaged by Airyscan super-resolution microscopy. Left: maximum intensity-projection images of Z stacks; middle: single Z-plane of images; right: co-localization (yellow) was quantified by determining the percentages of GFP-Rab5-positive (green) puncta containing mCherry-Rab7 (red) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. The object-based co-localization macro tool FIJI was employed. (C) HeLa cells stably expressing control or CPAP-shRNA were transfected with control vector (GFP), GFP-Rab7 (WT; wild-type) or GFP-Rab7 (DN; dominant negative) vector constructs, treated with AF555-EGF ligand for 60 min, and stained for CD63 to mark MVBs/late endosomes, and imaged by confocal microscopy to determine AF555-EGF and CD63 co-localization. Left top row: GFP expression in control and Rab7 construct-expressing cells; left bottom rows: representative single Z-plane of images showing ligand-bound EGFR-CD63 co-localization; right: co-localization (yellow) was quantified by determining the percentages of EGF-positive (red) puncta containing CD63 (green) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. Zoomed images correspond to the dashed inset boxes of the indicated images. Scale bars: 10 μm. p values: ∗∗∗∗<0.0001 by unpaired nonparametric Mann-Whitney test.

Techniques Used: Expressing, Control, shRNA, Staining, Super-Resolution Microscopy, Transfection, Construct, Stable Transfection, Plasmid Preparation, Dominant Negative Mutation, Confocal Microscopy, MANN-WHITNEY

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Journal: iScience

Article Title: Centrosomal P4.1-associated protein is a novel regulator of ESCRT pathway function during endosome maturation

doi: 10.1016/j.isci.2026.114659

Figure Lengend Snippet: CPAP depletion disrupts Rab5-to-Rab7 conversion (A) HeLa cells expressing control or CPAP shRNA were treated with untagged EGF for 60 min, stained for Rab5 and Rab7, and imaged by Airyscan super-resolution microscopy. Left: maximum intensity-projection images of Z stacks; middle: single Z-plane of images; right: co-localization (yellow) was quantified by determining the percentages of Rab5-positive (green) puncta containing Rab7 (red) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. (B) HeLa cells expressing control or CPAP shRNA were transfected with GFP-Rab5 and mCherry-Rab7 constructs, treated with untagged EGF for 60 min, and imaged by Airyscan super-resolution microscopy. Left: maximum intensity-projection images of Z stacks; middle: single Z-plane of images; right: co-localization (yellow) was quantified by determining the percentages of GFP-Rab5-positive (green) puncta containing mCherry-Rab7 (red) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. The object-based co-localization macro tool FIJI was employed. (C) HeLa cells stably expressing control or CPAP-shRNA were transfected with control vector (GFP), GFP-Rab7 (WT; wild-type) or GFP-Rab7 (DN; dominant negative) vector constructs, treated with AF555-EGF ligand for 60 min, and stained for CD63 to mark MVBs/late endosomes, and imaged by confocal microscopy to determine AF555-EGF and CD63 co-localization. Left top row: GFP expression in control and Rab7 construct-expressing cells; left bottom rows: representative single Z-plane of images showing ligand-bound EGFR-CD63 co-localization; right: co-localization (yellow) was quantified by determining the percentages of EGF-positive (red) puncta containing CD63 (green) puncta in representative single Z-planes of each cell and quantified from multiple cells across at least three experiments. Zoomed images correspond to the dashed inset boxes of the indicated images. Scale bars: 10 μm. p values: ∗∗∗∗<0.0001 by unpaired nonparametric Mann-Whitney test.

Article Snippet: GFP-Rab5 DN plasmid , Addgene , 28045.

Techniques: Expressing, Control, shRNA, Staining, Super-Resolution Microscopy, Transfection, Construct, Stable Transfection, Plasmid Preparation, Dominant Negative Mutation, Confocal Microscopy, MANN-WHITNEY

CTLA‐4 colocalizes with endosomal Rab GTPases. (a,b) CTLA‐4‐transduced HeLa (a) or Jurkat (b) cells were fixed, permeabilized, and stained with human anti‐CTLA‐4 Ab and rabbit anti‐Rab5, anti‐Rab7, anti‐Rab9 or anti‐Rab11 Abs followed by goat anti‐human IgG‐Alexa Fluor 546, donkey anti‐rabbit IgG‐Alexa Fluor 488, Hoechst and CTV. Cells were analysed by confocal microscopy. Scale bars = 10 μm. (c) Graph showing quantification of the colocalization of CTLA‐4 vesicles with Rab vesicles in (a) or (b). Data shown as mean ± SEM, n = 3. *** P < 0.005 determined by two‐way ANOVA with Sidak's multiple comparisons test

Journal: Immunology

Article Title: Regulation of CTLA‐4 recycling by LRBA and Rab11

doi: 10.1111/imm.13343

Figure Lengend Snippet: CTLA‐4 colocalizes with endosomal Rab GTPases. (a,b) CTLA‐4‐transduced HeLa (a) or Jurkat (b) cells were fixed, permeabilized, and stained with human anti‐CTLA‐4 Ab and rabbit anti‐Rab5, anti‐Rab7, anti‐Rab9 or anti‐Rab11 Abs followed by goat anti‐human IgG‐Alexa Fluor 546, donkey anti‐rabbit IgG‐Alexa Fluor 488, Hoechst and CTV. Cells were analysed by confocal microscopy. Scale bars = 10 μm. (c) Graph showing quantification of the colocalization of CTLA‐4 vesicles with Rab vesicles in (a) or (b). Data shown as mean ± SEM, n = 3. *** P < 0.005 determined by two‐way ANOVA with Sidak's multiple comparisons test

Article Snippet: The following Rab‐GFP constructs were acquired from Addgene and generated by Marci Scidmore (Rzomp et al; Cortes et al) (Rab4 WT, DN, CA; Rab5 WT; Rab11 CA), Sergio Grinstein (Bohdanowicz et al) (Rab5 DN, CA) and Richard Pagano (Choudhury et al) (Rab9 WT, DN; Rab11 WT, DN): Rab4WT (Addgene plasmid #49434), Rab4 DN (#49476), Rab4 CA (#49475), Rab5 WT (#49888), Rab5 DN (#35141), Rab5 CA (#35140), Rab9 WT (#12663), Rab9 DN (#12664), Rab11 WT (# 12674), Rab11 DN (#12678) and Rab11 CA (#49553).

Techniques: Staining, Confocal Microscopy

CTLA‐4 surface expression is regulated by Rab5 and Rab11. CTLA‐4‐transduced HeLa or Jurkat cells were transfected with Rab‐GFP constructs (empty vector—EV; wild type—WT; dominant negative—DN; or constitutively active—CA) for 24 h, and then stained for surface CTLA‐4 expression. (a) Flow cytometry contour plots of HeLa cells showing Rab‐GFP vs surface CTLA‐4 expression. (b) Flow cytometry plot showing the GFP + gate used. (c,d) Graphs showing surface CTLA‐4 expression relative to empty vector control in GFP + gated HeLa (c) or Jurkat (d) cells. Data shown as mean ± SD, c: n ≥ 3, d: n ≥ 4. Differences between conditions and EV determined by one‐way ANOVA and Dunnett's multiple comparisons test; * P < 0.05, *** P < 0.005 and **** P < 0.001

Journal: Immunology

Article Title: Regulation of CTLA‐4 recycling by LRBA and Rab11

doi: 10.1111/imm.13343

Figure Lengend Snippet: CTLA‐4 surface expression is regulated by Rab5 and Rab11. CTLA‐4‐transduced HeLa or Jurkat cells were transfected with Rab‐GFP constructs (empty vector—EV; wild type—WT; dominant negative—DN; or constitutively active—CA) for 24 h, and then stained for surface CTLA‐4 expression. (a) Flow cytometry contour plots of HeLa cells showing Rab‐GFP vs surface CTLA‐4 expression. (b) Flow cytometry plot showing the GFP + gate used. (c,d) Graphs showing surface CTLA‐4 expression relative to empty vector control in GFP + gated HeLa (c) or Jurkat (d) cells. Data shown as mean ± SD, c: n ≥ 3, d: n ≥ 4. Differences between conditions and EV determined by one‐way ANOVA and Dunnett's multiple comparisons test; * P < 0.05, *** P < 0.005 and **** P < 0.001

Techniques: Expressing, Transfection, Construct, Plasmid Preparation, Dominant Negative Mutation, Staining, Flow Cytometry

CTLA‐4 degradation is regulated by Rab5 and Rab7. CTLA‐4‐transduced HeLa or Jurkat cells were transfected with Rab‐GFP constructs (empty vector—EV; wild type—WT; dominant negative—DN; or constitutively active—CA) for 24 h, and then stained with anti‐CTLA‐4 PE at 37°C for 1 h. Cells were then incubated at 37°C for up to 4 h, before being fixed, and analysed by flow cytometry. (a,c) Flow cytometry histogram of HeLa (a) or Jurkat (c) cells showing GFP + gate used for analysis. (b,d) Graphs showing CTLA‐4 staining in GFP + gated HeLa (b) or Jurkat (d) cells, relative to 0 h. Data shown as mean ± SD, n ≥ 3. Differences between conditions were determined by two‐way ANOVA and Dunnett's multiple comparisons test; * P < 0.05, ** P < 0.01 and **** P < 0.001, ns—non‐significant

Journal: Immunology

Article Title: Regulation of CTLA‐4 recycling by LRBA and Rab11

doi: 10.1111/imm.13343

Figure Lengend Snippet: CTLA‐4 degradation is regulated by Rab5 and Rab7. CTLA‐4‐transduced HeLa or Jurkat cells were transfected with Rab‐GFP constructs (empty vector—EV; wild type—WT; dominant negative—DN; or constitutively active—CA) for 24 h, and then stained with anti‐CTLA‐4 PE at 37°C for 1 h. Cells were then incubated at 37°C for up to 4 h, before being fixed, and analysed by flow cytometry. (a,c) Flow cytometry histogram of HeLa (a) or Jurkat (c) cells showing GFP + gate used for analysis. (b,d) Graphs showing CTLA‐4 staining in GFP + gated HeLa (b) or Jurkat (d) cells, relative to 0 h. Data shown as mean ± SD, n ≥ 3. Differences between conditions were determined by two‐way ANOVA and Dunnett's multiple comparisons test; * P < 0.05, ** P < 0.01 and **** P < 0.001, ns—non‐significant

Techniques: Transfection, Construct, Plasmid Preparation, Dominant Negative Mutation, Staining, Incubation, Flow Cytometry

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1) Product Images from "Centrosomal P4.1-associated protein is a novel regulator of ESCRT pathway function during endosome maturation"

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