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app695 aenata (Addgene inc)

Name: app695 aenata
Brand: Addgene inc
Rating: 93 (2 reviews)

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Addgene inc app695 aenata

N2a cells transfected with <t>APP695</t> and LAMP1-mCh (red) and incubated with fluorescently-tagged A) N-terminal anti-APP (APP), B) Aβ-region targeting 6E10 antibodies, or C) anti-NCAM antibodies (green) on ice for 20 minutes, then immediately fixed or allowed to incubate at 37°C for 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). D) Quantification of the mean % of APP colocalized with LAMP1 from three replicate experiments (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. E) N2a cells transfected with APP695 and LAMP1-mCh (red) that were treated with DMSO, 20μM Pitstop2 or 10μM EIPA. APP was then bound/crosslinked by antibody and imaged after 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). F) Quantification of the mean % of APP colocalized with LAMP1 (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. Data is presented as mean ± SEM. * p<0.05; Scale bar = 5μm .

App695 Aenata, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA"

Article Title: Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA

Journal: bioRxiv

doi: 10.1101/2025.03.02.641070

N2a cells transfected with APP695 and LAMP1-mCh (red) and incubated with fluorescently-tagged A) N-terminal anti-APP (APP), B) Aβ-region targeting 6E10 antibodies, or C) anti-NCAM antibodies (green) on ice for 20 minutes, then immediately fixed or allowed to incubate at 37°C for 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). D) Quantification of the mean % of APP colocalized with LAMP1 from three replicate experiments (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. E) N2a cells transfected with APP695 and LAMP1-mCh (red) that were treated with DMSO, 20μM Pitstop2 or 10μM EIPA. APP was then bound/crosslinked by antibody and imaged after 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). F) Quantification of the mean % of APP colocalized with LAMP1 (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. Data is presented as mean ± SEM. * p<0.05; Scale bar = 5μm .

Figure Legend Snippet: N2a cells transfected with APP695 and LAMP1-mCh (red) and incubated with fluorescently-tagged A) N-terminal anti-APP (APP), B) Aβ-region targeting 6E10 antibodies, or C) anti-NCAM antibodies (green) on ice for 20 minutes, then immediately fixed or allowed to incubate at 37°C for 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). D) Quantification of the mean % of APP colocalized with LAMP1 from three replicate experiments (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. E) N2a cells transfected with APP695 and LAMP1-mCh (red) that were treated with DMSO, 20μM Pitstop2 or 10μM EIPA. APP was then bound/crosslinked by antibody and imaged after 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). F) Quantification of the mean % of APP colocalized with LAMP1 (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. Data is presented as mean ± SEM. * p<0.05; Scale bar = 5μm .

Techniques Used: Transfection, Incubation

A) N2a cells transfected with Fe65-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were incubated with fluorescent anti-APP or anti-NCAM antibodies (red) on ice, then immediately fixed on ice as a baseline (00:00) or incubated for 30 seconds (00:30), 2 minutes (02:00), 5 minutes (5:00), and 10 minutes (10:00). Colocalization between signals is indicated by white pixels. B/C) Quantification of colocalization data (n=3; 15 images per replicate) is represented as the difference between the colocalization at each timepoint to the colocalization at baseline (mean % colocalized at XX:XX – mean % colocalized at 00:00). * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. D) N2a cells transfected with Arf6-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment above was repeated with Arf6-EGFP expressing cells. Colocalization between signals is indicated by white pixels. E/F) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from G/H) was repeated for N2a cells expressing Arf6-GFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. Significance was measured by a two-way ANOVA with a Tukey’s test using a single pooled variance. Data is presented as mean ± SEM. */# p<0.05; Scale bar = 5μm .

Figure Legend Snippet: A) N2a cells transfected with Fe65-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were incubated with fluorescent anti-APP or anti-NCAM antibodies (red) on ice, then immediately fixed on ice as a baseline (00:00) or incubated for 30 seconds (00:30), 2 minutes (02:00), 5 minutes (5:00), and 10 minutes (10:00). Colocalization between signals is indicated by white pixels. B/C) Quantification of colocalization data (n=3; 15 images per replicate) is represented as the difference between the colocalization at each timepoint to the colocalization at baseline (mean % colocalized at XX:XX – mean % colocalized at 00:00). * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. D) N2a cells transfected with Arf6-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment above was repeated with Arf6-EGFP expressing cells. Colocalization between signals is indicated by white pixels. E/F) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from G/H) was repeated for N2a cells expressing Arf6-GFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. Significance was measured by a two-way ANOVA with a Tukey’s test using a single pooled variance. Data is presented as mean ± SEM. */# p<0.05; Scale bar = 5μm .

Techniques Used: Transfection, Incubation, Expressing

A) N2a cells transfected with Rac1-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were incubated with tagged anti-APP or anti-NCAM antibodies (red) on ice and then were either immediately fixed on ice as a baseline (00:00), or incubated for 30 seconds (00:30), 2 minutes (02:00), 5 minutes (5:00), and 10 minutes (10:00) at 37°C 5% CO2 prior to fixation. Colocalization between signals is indicated by white pixels. B/C) Quantification of colocalization data (n=3; 15 images per replicate) is represented as the difference between the colocalization at each timepoint to the colocalization at baseline (mean % colocalized at XX:XX – mean % colocalized at 00:00). * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. D) N2a cells transfected with Cdc42-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment from A) was repeated with Cdc42-EGFP expressing cells. Colocalization between signals is indicated by white pixels. E/F) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from B/C) was repeated for N2a cells expressing Cdc42-EGFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. G) N2a cells transfected with RhoA-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment from A) was repeated with RhoA-EGFP expressing cells. Colocalization between signals is indicated by white pixels. H/I) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from B/C) was repeated for N2a cells expressing RhoA-EGFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. Significance was measured by a two-way ANOVA with a Tukey’s test using a single pooled variance. Data is presented as mean ± SEM. */# p<0.05; Scale bar = 5μm .

Figure Legend Snippet: A) N2a cells transfected with Rac1-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were incubated with tagged anti-APP or anti-NCAM antibodies (red) on ice and then were either immediately fixed on ice as a baseline (00:00), or incubated for 30 seconds (00:30), 2 minutes (02:00), 5 minutes (5:00), and 10 minutes (10:00) at 37°C 5% CO2 prior to fixation. Colocalization between signals is indicated by white pixels. B/C) Quantification of colocalization data (n=3; 15 images per replicate) is represented as the difference between the colocalization at each timepoint to the colocalization at baseline (mean % colocalized at XX:XX – mean % colocalized at 00:00). * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. D) N2a cells transfected with Cdc42-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment from A) was repeated with Cdc42-EGFP expressing cells. Colocalization between signals is indicated by white pixels. E/F) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from B/C) was repeated for N2a cells expressing Cdc42-EGFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. G) N2a cells transfected with RhoA-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment from A) was repeated with RhoA-EGFP expressing cells. Colocalization between signals is indicated by white pixels. H/I) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from B/C) was repeated for N2a cells expressing RhoA-EGFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. Significance was measured by a two-way ANOVA with a Tukey’s test using a single pooled variance. Data is presented as mean ± SEM. */# p<0.05; Scale bar = 5μm .

Techniques Used: Transfection, Incubation, Expressing

A) N2a cells transfected with Fe65-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following an incubation for 30 seconds. Colocalization was assessed between Fe65 and antibody bound/crosslinked APP or PLC8PH (white pixels). B) Quantification of the mean % of Fe65 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). C) N2a cells transfected with Arf6-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following a 30 second incubation. Colocalization was assessed between Arf6 and bound/crosslinked APP or PLC8PH (white pixels). D) Quantification of the mean % of Arf6 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). Significant changes in the percentage colocalized between treatments were assessed by a one-way ANOVA with Tukey’s test . Data is presented as mean ± SEM. *p<0.05; Scale bar = 5μm .

Figure Legend Snippet: A) N2a cells transfected with Fe65-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following an incubation for 30 seconds. Colocalization was assessed between Fe65 and antibody bound/crosslinked APP or PLC8PH (white pixels). B) Quantification of the mean % of Fe65 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). C) N2a cells transfected with Arf6-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following a 30 second incubation. Colocalization was assessed between Arf6 and bound/crosslinked APP or PLC8PH (white pixels). D) Quantification of the mean % of Arf6 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). Significant changes in the percentage colocalized between treatments were assessed by a one-way ANOVA with Tukey’s test . Data is presented as mean ± SEM. *p<0.05; Scale bar = 5μm .

Techniques Used: Transfection, Incubation

A) N2a cells transfected with Rac1-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following a 30 second incubation. Colocalization was assessed between Rac1 and bound/crosslinked APP or PLC8PH (white pixels). B) Quantification of the mean % of Rac1 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). C) N2a cells transfected with Cdc42-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated as described in A), then incubated with tagged anti-APP antibodies (red) on ice and fixed following a 30 second incubation. Colocalization was assessed between Cdc42 and bound/crosslinked APP or PLC8PH (white pixels). D) Quantification of the mean % of Cdc42 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). E) N2a cells transfected with RhoA-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated as described in A), then incubated with tagged anti-APP antibodies (red) on ice and then fixed following an incubation for 30 seconds. Colocalization was assessed between RhoA and bound/crosslinked APP or PLC8PH (white pixels). F) Quantification of the mean % of RhoA colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). Significant changes in the percentage colocalized between treatments were assessed by a one-way ANOVA with Tukey’s test . Data is presented as mean ± SEM. *p<0.05; Scale bar = 5μm .

Figure Legend Snippet: A) N2a cells transfected with Rac1-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following a 30 second incubation. Colocalization was assessed between Rac1 and bound/crosslinked APP or PLC8PH (white pixels). B) Quantification of the mean % of Rac1 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). C) N2a cells transfected with Cdc42-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated as described in A), then incubated with tagged anti-APP antibodies (red) on ice and fixed following a 30 second incubation. Colocalization was assessed between Cdc42 and bound/crosslinked APP or PLC8PH (white pixels). D) Quantification of the mean % of Cdc42 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). E) N2a cells transfected with RhoA-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated as described in A), then incubated with tagged anti-APP antibodies (red) on ice and then fixed following an incubation for 30 seconds. Colocalization was assessed between RhoA and bound/crosslinked APP or PLC8PH (white pixels). F) Quantification of the mean % of RhoA colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). Significant changes in the percentage colocalized between treatments were assessed by a one-way ANOVA with Tukey’s test . Data is presented as mean ± SEM. *p<0.05; Scale bar = 5μm .

Techniques Used: Transfection, Incubation

Image Search Results

Journal: bioRxiv

Article Title: Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA

doi: 10.1101/2025.03.02.641070

Figure Lengend Snippet: N2a cells transfected with APP695 and LAMP1-mCh (red) and incubated with fluorescently-tagged A) N-terminal anti-APP (APP), B) Aβ-region targeting 6E10 antibodies, or C) anti-NCAM antibodies (green) on ice for 20 minutes, then immediately fixed or allowed to incubate at 37°C for 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). D) Quantification of the mean % of APP colocalized with LAMP1 from three replicate experiments (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. E) N2a cells transfected with APP695 and LAMP1-mCh (red) that were treated with DMSO, 20μM Pitstop2 or 10μM EIPA. APP was then bound/crosslinked by antibody and imaged after 15 minutes. Colocalization was assessed between crosslinked APP and LAMP1 (white pixels). F) Quantification of the mean % of APP colocalized with LAMP1 (n=3; 10 images per replicate), with significance calculated by a one-way ANOVA with Tukey’s test. Data is presented as mean ± SEM. * p<0.05; Scale bar = 5μm .

Article Snippet: APP695-AENATA (pCAX-APP-AENATA) was a gift from Dennis Selkoe and Tracy Young-Pearse (Addgene plasmid #30144).

Techniques: Transfection, Incubation

Journal: bioRxiv

Article Title: Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA

doi: 10.1101/2025.03.02.641070

Figure Lengend Snippet: A) N2a cells transfected with Fe65-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were incubated with fluorescent anti-APP or anti-NCAM antibodies (red) on ice, then immediately fixed on ice as a baseline (00:00) or incubated for 30 seconds (00:30), 2 minutes (02:00), 5 minutes (5:00), and 10 minutes (10:00). Colocalization between signals is indicated by white pixels. B/C) Quantification of colocalization data (n=3; 15 images per replicate) is represented as the difference between the colocalization at each timepoint to the colocalization at baseline (mean % colocalized at XX:XX – mean % colocalized at 00:00). * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. D) N2a cells transfected with Arf6-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment above was repeated with Arf6-EGFP expressing cells. Colocalization between signals is indicated by white pixels. E/F) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from G/H) was repeated for N2a cells expressing Arf6-GFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. Significance was measured by a two-way ANOVA with a Tukey’s test using a single pooled variance. Data is presented as mean ± SEM. */# p<0.05; Scale bar = 5μm .

Article Snippet: APP695-AENATA (pCAX-APP-AENATA) was a gift from Dennis Selkoe and Tracy Young-Pearse (Addgene plasmid #30144).

Techniques: Transfection, Incubation, Expressing

Journal: bioRxiv

Article Title: Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA

doi: 10.1101/2025.03.02.641070

Figure Lengend Snippet: A) N2a cells transfected with Rac1-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were incubated with tagged anti-APP or anti-NCAM antibodies (red) on ice and then were either immediately fixed on ice as a baseline (00:00), or incubated for 30 seconds (00:30), 2 minutes (02:00), 5 minutes (5:00), and 10 minutes (10:00) at 37°C 5% CO2 prior to fixation. Colocalization between signals is indicated by white pixels. B/C) Quantification of colocalization data (n=3; 15 images per replicate) is represented as the difference between the colocalization at each timepoint to the colocalization at baseline (mean % colocalized at XX:XX – mean % colocalized at 00:00). * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. D) N2a cells transfected with Cdc42-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment from A) was repeated with Cdc42-EGFP expressing cells. Colocalization between signals is indicated by white pixels. E/F) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from B/C) was repeated for N2a cells expressing Cdc42-EGFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. G) N2a cells transfected with RhoA-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Experiment from A) was repeated with RhoA-EGFP expressing cells. Colocalization between signals is indicated by white pixels. H/I) Quantification of colocalization data (n=3; 15 images per replicate). Data representation and analysis from B/C) was repeated for N2a cells expressing RhoA-EGFP. * denotes significant difference to baseline within condition; # denotes significant difference between groups at the indicated timepoint. Significance was measured by a two-way ANOVA with a Tukey’s test using a single pooled variance. Data is presented as mean ± SEM. */# p<0.05; Scale bar = 5μm .

Article Snippet: APP695-AENATA (pCAX-APP-AENATA) was a gift from Dennis Selkoe and Tracy Young-Pearse (Addgene plasmid #30144).

Techniques: Transfection, Incubation, Expressing

Journal: bioRxiv

Article Title: Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA

doi: 10.1101/2025.03.02.641070

Figure Lengend Snippet: A) N2a cells transfected with Fe65-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following an incubation for 30 seconds. Colocalization was assessed between Fe65 and antibody bound/crosslinked APP or PLC8PH (white pixels). B) Quantification of the mean % of Fe65 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). C) N2a cells transfected with Arf6-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following a 30 second incubation. Colocalization was assessed between Arf6 and bound/crosslinked APP or PLC8PH (white pixels). D) Quantification of the mean % of Arf6 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). Significant changes in the percentage colocalized between treatments were assessed by a one-way ANOVA with Tukey’s test . Data is presented as mean ± SEM. *p<0.05; Scale bar = 5μm .

Article Snippet: APP695-AENATA (pCAX-APP-AENATA) was a gift from Dennis Selkoe and Tracy Young-Pearse (Addgene plasmid #30144).

Techniques: Transfection, Incubation

Journal: bioRxiv

Article Title: Macropinocytosis of amyloid precursor protein requires the adaptor protein Fe65 and the recruitment and activity of Arf6 and the RhoGTPases Rac1, Cdc42 and RhoA

doi: 10.1101/2025.03.02.641070

Figure Lengend Snippet: A) N2a cells transfected with Rac1-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated with 0.1% DMSO, 5μM NAV-2729, 10μM EHT 1864, 10μM ML 141, and 35μM Rhosin. After treatment, cells were incubated with tagged anti-APP antibodies (red) on ice and then fixed following a 30 second incubation. Colocalization was assessed between Rac1 and bound/crosslinked APP or PLC8PH (white pixels). B) Quantification of the mean % of Rac1 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). C) N2a cells transfected with Cdc42-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated as described in A), then incubated with tagged anti-APP antibodies (red) on ice and fixed following a 30 second incubation. Colocalization was assessed between Cdc42 and bound/crosslinked APP or PLC8PH (white pixels). D) Quantification of the mean % of Cdc42 colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). E) N2a cells transfected with RhoA-EGFP (green), PLC8PH-mRFP (magenta) and APP695. Cells were treated as described in A), then incubated with tagged anti-APP antibodies (red) on ice and then fixed following an incubation for 30 seconds. Colocalization was assessed between RhoA and bound/crosslinked APP or PLC8PH (white pixels). F) Quantification of the mean % of RhoA colocalized with APP (left) or PLC8PH (right) from three replicate experiments (n=3; 15 images per replicate). Significant changes in the percentage colocalized between treatments were assessed by a one-way ANOVA with Tukey’s test . Data is presented as mean ± SEM. *p<0.05; Scale bar = 5μm .

Article Snippet: APP695-AENATA (pCAX-APP-AENATA) was a gift from Dennis Selkoe and Tracy Young-Pearse (Addgene plasmid #30144).

Techniques: Transfection, Incubation

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