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3d image processing software imaris  (Oxford Instruments)
99
Oxford Instruments 3d image processing software imaris
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
3d Image Processing Software Imaris, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fnirs system data processing software  (Shimadzu Corporation)
99
Shimadzu Corporation fnirs system data processing software
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
Fnirs System Data Processing Software, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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avizo image processing software  (Thermo Fisher)
90
Thermo Fisher avizo image processing software
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
Avizo Image Processing Software, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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3-d post-processing software cardiacct syngoviatm  (Siemens Healthineers)
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Siemens Healthineers 3-d post-processing software cardiacct syngoviatm
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
3 D Post Processing Software Cardiacct Syngoviatm, supplied by Siemens Healthineers, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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image processing software oxford instruments  (Oxford Instruments)
99
Oxford Instruments image processing software oxford instruments
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
Image Processing Software Oxford Instruments, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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image processing software imaris  (Oxford Instruments)
99
Oxford Instruments image processing software imaris
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
Image Processing Software Imaris, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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data processing software  (Inscopix Inc)
90
Inscopix Inc data processing software
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
Data Processing Software, supplied by Inscopix Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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data processing software (idps)  (Inscopix Inc)
90
Inscopix Inc data processing software (idps)
Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch <t>and</t> <t>SNAP-tag.</t> The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) <t>3D</t> maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM
Data Processing Software (Idps), supplied by Inscopix Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch and SNAP-tag. The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) 3D maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM

Journal: Journal of Translational Medicine

Article Title: Detection of extracellular amyloid beta aggregates by an Aducanumab-based synNotch receptor: an in vitro proof-of-concept study

doi: 10.1186/s12967-025-07324-2

Figure Lengend Snippet: Assessment of Aβ-targeting Adu-synNotch activation in an in vitro model of Aβ . (a) Expression cassette of EF1A-regulated Adu-synNotch and SNAP-tag. The VLC and VHC of Aducanumab is fused by a flexible G4S(3) linker to form an scFv, with an N-terminal human CD8α signal peptide (SP; white) for membrane localization. The SNAP-tag has an NLS to target the reporter to the nucleus. Adu-synNotch and SNAP-tag are separated by a T2A sequence for bicistronic expression. (b) Gal4UASmCMV-regulated CLIP-tag under the control of Adu-synNotch activation. The CLIP-tag contains an NLS for nuclear targeting. (c) Adu-synNotch regulating CLIP-tag expression in NIH 3T3 cells. (1) Adu-synNotch and SNAP-tag are constitutively expressed. SNAP expression in the nucleus is proportional to synNotch transcription. (2) Activation of Adu-synNotch by Aβ. (3) Adu-synNotch activation results in the expression of nuclear-targeted CLIP-tag. (4) Adu-synNotch activation results in colocalization of SNAP-tag and CLIP-tag, used to normalize Adu-synNotch activation to copy-number integration. (d) Schematic overview of the experiments. A day after plating cells, AβO (1−42) treatments are added for 48 h, after which cells are labeled with CLIP-tag and SNAP-tag substrates and processed for confocal image analyses. (e) 3D maximum intensity projection confocal images of clones expressing Adu-synNotch under the indicated treatment conditions. Yellow boxes are an expanded view of regions within the tile scans. DAPI for cell nuclei, SNAP-tag for transduced cells, and CLIP-tag for synNotch activation. DAPI + /SNAP − white arrowheads (untransduced), DAPI + /SNAP + /CLIP − yellow arrows (inactive Adu-synNotch), DAPI + /SNAP + /CLIP + white arrows (active Adu-synNotch). Scale bar: 200 μm tile scans; 40 μm expanded view. (f-i) Quantification of (f) the number of SNAP-positive cells, (g) mean SNAP and (h) CLIP as well as (i) CLIP to SNAP ratio. Voxel intensity is in arbitrary units (au). Statistics: (f , h , i) Independent t -test and (g) Welch’s t -test. Two-tails. Three independent experiments. * p <.05, ** p <.01. Error Bars 1x SEM

Article Snippet: For the quantification and statistical analyses of Adu-synNotch activation, we generated three-dimensional regions of interest (ROIs) based on the SNAP channel using the 3D image processing software IMARIS.

Techniques: Activation Assay, In Vitro, Expressing, Membrane, Sequencing, Control, Labeling, Clone Assay