addgene usp19 er Search Results


addgene usp19 er  (Addgene inc)
93
Addgene inc addgene usp19 er
Identification of <t>USP19</t> as a TDP-43-directed deubiquitinase. ( A ) Schematic of Duolink® PLA principle for detection of ubiquitin (Ub)-TDP-43 complexes. ( B ) Representative images of Ub-TDP-43 PLA puncta (red), TDP-43 (green), and DAPI (blue) in tetracycline (tet)-induced HeLa-GFP-TDP-43 cells transfected with Smartpool siRNAs targeting 20 different USPs. ( C ) Quantification of Ub-TDP-43 PLA puncta numbers per cell (one-way ANOVA, F(20, 42) = 2.711, P = 0.0032; post hoc Dunnett compared to control siRNA (Cont), * P = 0.0270; n = 3 independent experiments per USP with 4 to 9 images averaged per experiment). ( D ) Schematic of USP19-flag constructs: ER-localized USP19 <t>(USP19-ER),</t> cytosolic USP19 (USP19-Cyto), and USP19-ER lacking catalytic DUB activity (USP19-CS-ER). ( E ) Representative blots showing co-IP of Myc-TDP-43 with USP19-flag variants (flag pulldown) in HEK293T cells cotransfected ± USP19-flag variants or vector control ± Myc-TDP-43. ( F ) Representative images of USP19–TDP-43 PLA puncta (red: myc + flag PLA) and DAPI (blue) from HeLa cells transfected with Myc-TDP-43 and USP19-flag variants or vector control. One probe - lacks one secondary antibody probe. ( G ) Representative blots of polyubiquitin conjugates (Ub-K48 & Ub-K63) in TDP-43 pulldown (myc) from HEK293T cells transfected with ubiquitin-HA (Ub-HA) and myc-TDP-43 with USP19-flag variants or vector control.
Addgene Usp19 Er, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/addgene+usp19+er/pmc12994178-156-6-6?v=Addgene+inc
Average 93 stars, based on 1 article reviews
addgene usp19 er - by Bioz Stars, 2026-08
93/100 stars
  Buy from Supplier
plasmids 78597  (Addgene inc)
93
Addgene inc plasmids 78597
Identification of <t>USP19</t> as a TDP-43-directed deubiquitinase. ( A ) Schematic of Duolink® PLA principle for detection of ubiquitin (Ub)-TDP-43 complexes. ( B ) Representative images of Ub-TDP-43 PLA puncta (red), TDP-43 (green), and DAPI (blue) in tetracycline (tet)-induced HeLa-GFP-TDP-43 cells transfected with Smartpool siRNAs targeting 20 different USPs. ( C ) Quantification of Ub-TDP-43 PLA puncta numbers per cell (one-way ANOVA, F(20, 42) = 2.711, P = 0.0032; post hoc Dunnett compared to control siRNA (Cont), * P = 0.0270; n = 3 independent experiments per USP with 4 to 9 images averaged per experiment). ( D ) Schematic of USP19-flag constructs: ER-localized USP19 <t>(USP19-ER),</t> cytosolic USP19 (USP19-Cyto), and USP19-ER lacking catalytic DUB activity (USP19-CS-ER). ( E ) Representative blots showing co-IP of Myc-TDP-43 with USP19-flag variants (flag pulldown) in HEK293T cells cotransfected ± USP19-flag variants or vector control ± Myc-TDP-43. ( F ) Representative images of USP19–TDP-43 PLA puncta (red: myc + flag PLA) and DAPI (blue) from HeLa cells transfected with Myc-TDP-43 and USP19-flag variants or vector control. One probe - lacks one secondary antibody probe. ( G ) Representative blots of polyubiquitin conjugates (Ub-K48 & Ub-K63) in TDP-43 pulldown (myc) from HEK293T cells transfected with ubiquitin-HA (Ub-HA) and myc-TDP-43 with USP19-flag variants or vector control.
Plasmids 78597, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/addgene+usp19+er/pmc09842591-42-19-18?v=Addgene+inc
Average 93 stars, based on 1 article reviews
plasmids 78597 - by Bioz Stars, 2026-08
93/100 stars
  Buy from Supplier

Image Search Results


Identification of USP19 as a TDP-43-directed deubiquitinase. ( A ) Schematic of Duolink® PLA principle for detection of ubiquitin (Ub)-TDP-43 complexes. ( B ) Representative images of Ub-TDP-43 PLA puncta (red), TDP-43 (green), and DAPI (blue) in tetracycline (tet)-induced HeLa-GFP-TDP-43 cells transfected with Smartpool siRNAs targeting 20 different USPs. ( C ) Quantification of Ub-TDP-43 PLA puncta numbers per cell (one-way ANOVA, F(20, 42) = 2.711, P = 0.0032; post hoc Dunnett compared to control siRNA (Cont), * P = 0.0270; n = 3 independent experiments per USP with 4 to 9 images averaged per experiment). ( D ) Schematic of USP19-flag constructs: ER-localized USP19 (USP19-ER), cytosolic USP19 (USP19-Cyto), and USP19-ER lacking catalytic DUB activity (USP19-CS-ER). ( E ) Representative blots showing co-IP of Myc-TDP-43 with USP19-flag variants (flag pulldown) in HEK293T cells cotransfected ± USP19-flag variants or vector control ± Myc-TDP-43. ( F ) Representative images of USP19–TDP-43 PLA puncta (red: myc + flag PLA) and DAPI (blue) from HeLa cells transfected with Myc-TDP-43 and USP19-flag variants or vector control. One probe - lacks one secondary antibody probe. ( G ) Representative blots of polyubiquitin conjugates (Ub-K48 & Ub-K63) in TDP-43 pulldown (myc) from HEK293T cells transfected with ubiquitin-HA (Ub-HA) and myc-TDP-43 with USP19-flag variants or vector control.

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Ubiquitin-specific peptidase-19 links TDP-43 aggregation to ER stress

doi: 10.1073/pnas.2514355123

Figure Lengend Snippet: Identification of USP19 as a TDP-43-directed deubiquitinase. ( A ) Schematic of Duolink® PLA principle for detection of ubiquitin (Ub)-TDP-43 complexes. ( B ) Representative images of Ub-TDP-43 PLA puncta (red), TDP-43 (green), and DAPI (blue) in tetracycline (tet)-induced HeLa-GFP-TDP-43 cells transfected with Smartpool siRNAs targeting 20 different USPs. ( C ) Quantification of Ub-TDP-43 PLA puncta numbers per cell (one-way ANOVA, F(20, 42) = 2.711, P = 0.0032; post hoc Dunnett compared to control siRNA (Cont), * P = 0.0270; n = 3 independent experiments per USP with 4 to 9 images averaged per experiment). ( D ) Schematic of USP19-flag constructs: ER-localized USP19 (USP19-ER), cytosolic USP19 (USP19-Cyto), and USP19-ER lacking catalytic DUB activity (USP19-CS-ER). ( E ) Representative blots showing co-IP of Myc-TDP-43 with USP19-flag variants (flag pulldown) in HEK293T cells cotransfected ± USP19-flag variants or vector control ± Myc-TDP-43. ( F ) Representative images of USP19–TDP-43 PLA puncta (red: myc + flag PLA) and DAPI (blue) from HeLa cells transfected with Myc-TDP-43 and USP19-flag variants or vector control. One probe - lacks one secondary antibody probe. ( G ) Representative blots of polyubiquitin conjugates (Ub-K48 & Ub-K63) in TDP-43 pulldown (myc) from HEK293T cells transfected with ubiquitin-HA (Ub-HA) and myc-TDP-43 with USP19-flag variants or vector control.

Article Snippet: The following plasmids were obtained from addgene: USP19-ER (#78597), USP19-Cyto (#78579), USP19-CS-ER (#78584), HA-Ubiquitin (#18712), USP19-ER-mCitrine (#78593), USP19-Cyto-mCitrine (#78595), USP19-CS-mCitrine (#78594), TDP-43-tomato-HA (#28205), and Cry2olig-mCherry (#60032). pcDNA (Invitrogen, Cat# V79020) was purchased from Invitrogen.

Techniques: Ubiquitin Proteomics, Transfection, Control, Construct, Activity Assay, Co-Immunoprecipitation Assay, Plasmid Preparation

USP19-ER isoform preferentially deubiquitinates and increases insoluble TDP-CTF through phase separation. ( A ) Representative blots of RIPA-soluble and RIPA-insoluble proteins from tet-induced GFP-TDP-43 cells transfected with vector control or USP19 variants. ( B – G ) Quantification of TDP-80 (GFP-TDP-43), TDP-43, and TDP-35 from RIPA-soluble (sol) and RIPA-insoluble (insol) fractions (Insol TDP-35: one-way ANOVA, F(3, 8) = 6.108, P = 0.0183; post hoc Dunnett, ** P < 0.01; ns: not significant; n = 3 independent experiments with 1 to 2 replicates averaged per experiment). ( H ) Representative blots of polyubiquitin conjugates (Ub-K48 & Ub-K63) in myc-TDP-35 pulldown (myc) from HEK293T cells transfected with ubiquitin-HA (Ub-HA) and myc-TDP-35 with USP19-flag variants or vector control. ( I ) Schematic model blue light-induced phase separation of Cry2olig-TDP-LCD-mCherry. ( J ) Representative images of Cry2olig-TDP-LCD-mCherry T(red) before and after blue light stimulation (488 nm, 1 s) in HeLa cells transfected with Cry2olig-TDP-LCD and USP19 variants (green) or vector control. ( K ) Quantification of Cry2olig-TDP-LCD puncta numbers normalized to cell area (two-way ANOVA, F(3, 39) = 21.33, P < 0.0001; post hoc Dunnett compared to control, **** P < 0.0001, ns: not significant; n = 2 independent experiments with 2 to 3 replicates averaged per experiment).

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Ubiquitin-specific peptidase-19 links TDP-43 aggregation to ER stress

doi: 10.1073/pnas.2514355123

Figure Lengend Snippet: USP19-ER isoform preferentially deubiquitinates and increases insoluble TDP-CTF through phase separation. ( A ) Representative blots of RIPA-soluble and RIPA-insoluble proteins from tet-induced GFP-TDP-43 cells transfected with vector control or USP19 variants. ( B – G ) Quantification of TDP-80 (GFP-TDP-43), TDP-43, and TDP-35 from RIPA-soluble (sol) and RIPA-insoluble (insol) fractions (Insol TDP-35: one-way ANOVA, F(3, 8) = 6.108, P = 0.0183; post hoc Dunnett, ** P < 0.01; ns: not significant; n = 3 independent experiments with 1 to 2 replicates averaged per experiment). ( H ) Representative blots of polyubiquitin conjugates (Ub-K48 & Ub-K63) in myc-TDP-35 pulldown (myc) from HEK293T cells transfected with ubiquitin-HA (Ub-HA) and myc-TDP-35 with USP19-flag variants or vector control. ( I ) Schematic model blue light-induced phase separation of Cry2olig-TDP-LCD-mCherry. ( J ) Representative images of Cry2olig-TDP-LCD-mCherry T(red) before and after blue light stimulation (488 nm, 1 s) in HeLa cells transfected with Cry2olig-TDP-LCD and USP19 variants (green) or vector control. ( K ) Quantification of Cry2olig-TDP-LCD puncta numbers normalized to cell area (two-way ANOVA, F(3, 39) = 21.33, P < 0.0001; post hoc Dunnett compared to control, **** P < 0.0001, ns: not significant; n = 2 independent experiments with 2 to 3 replicates averaged per experiment).

Article Snippet: The following plasmids were obtained from addgene: USP19-ER (#78597), USP19-Cyto (#78579), USP19-CS-ER (#78584), HA-Ubiquitin (#18712), USP19-ER-mCitrine (#78593), USP19-Cyto-mCitrine (#78595), USP19-CS-mCitrine (#78594), TDP-43-tomato-HA (#28205), and Cry2olig-mCherry (#60032). pcDNA (Invitrogen, Cat# V79020) was purchased from Invitrogen.

Techniques: Transfection, Plasmid Preparation, Control, Ubiquitin Proteomics