rabbit anti ankle2 Search Results


rabbit anti ankle2  (Cell Signaling Technology Inc)
93
Cell Signaling Technology Inc rabbit anti ankle2
Rabbit Anti Ankle2, supplied by Cell Signaling Technology 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/result/rabbit anti ankle2/product/Cell Signaling Technology Inc
Average 93 stars, based on 1 article reviews
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atlas antibodies iankle2 rabbit  (Atlas Antibodies)
92
Atlas Antibodies atlas antibodies iankle2 rabbit
Atlas Antibodies Iankle2 Rabbit, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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atlas antibodies iankle2 rabbit - by Bioz Stars, 2026-04
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anti-human ankle2 rabbit polyclonal  (Thermo Fisher)
90
Thermo Fisher anti-human ankle2 rabbit polyclonal
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Anti Human Ankle2 Rabbit Polyclonal, 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
https://www.bioz.com/result/anti-human ankle2 rabbit polyclonal/product/Thermo Fisher
Average 90 stars, based on 1 article reviews
anti-human ankle2 rabbit polyclonal - by Bioz Stars, 2026-04
90/100 stars
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rabbit anti-ankle2 ab225905  (Cell Signaling Technology Inc)
90
Cell Signaling Technology Inc rabbit anti-ankle2 ab225905
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Rabbit Anti Ankle2 Ab225905, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti-ankle2 ab225905/product/Cell Signaling Technology Inc
Average 90 stars, based on 1 article reviews
rabbit anti-ankle2 ab225905 - by Bioz Stars, 2026-04
90/100 stars
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rabbit anti ankle2  (Novus Biologicals)
93
Novus Biologicals rabbit anti ankle2
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Rabbit Anti Ankle2, supplied by Novus Biologicals, 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/result/rabbit anti ankle2/product/Novus Biologicals
Average 93 stars, based on 1 article reviews
rabbit anti ankle2 - by Bioz Stars, 2026-04
93/100 stars
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mouse anti-strep  (Qiagen)
90
Qiagen mouse anti-strep
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Mouse Anti Strep, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse anti-strep/product/Qiagen
Average 90 stars, based on 1 article reviews
mouse anti-strep - by Bioz Stars, 2026-04
90/100 stars
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anti lemd2  (Atlas Antibodies)
93
Atlas Antibodies anti lemd2
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Anti Lemd2, supplied by Atlas Antibodies, 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/result/anti lemd2/product/Atlas Antibodies
Average 93 stars, based on 1 article reviews
anti lemd2 - by Bioz Stars, 2026-04
93/100 stars
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rabbit anti chmp7  (Proteintech)
93
Proteintech rabbit anti chmp7
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Rabbit Anti Chmp7, supplied by Proteintech, 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/result/rabbit anti chmp7/product/Proteintech
Average 93 stars, based on 1 article reviews
rabbit anti chmp7 - by Bioz Stars, 2026-04
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anti tau phospho ser422 rabbit polyclonal  (GeneTex)
90
GeneTex anti tau phospho ser422 rabbit polyclonal
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Anti Tau Phospho Ser422 Rabbit Polyclonal, supplied by GeneTex, 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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Average 90 stars, based on 1 article reviews
anti tau phospho ser422 rabbit polyclonal - by Bioz Stars, 2026-04
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rabbit  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc rabbit
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Rabbit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit/product/Cell Signaling Technology Inc
Average 96 stars, based on 1 article reviews
rabbit - by Bioz Stars, 2026-04
96/100 stars
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rabbit polyclonal  (Abcam)
99
Abcam rabbit polyclonal
( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, <t>ANKLE2</t> and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.
Rabbit Polyclonal, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal/product/Abcam
Average 99 stars, based on 1 article reviews
rabbit polyclonal - by Bioz Stars, 2026-04
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Image Search Results


( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, ANKLE2 and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.

Journal: bioRxiv

Article Title: Genome-wide CRISPRi screening reveals regulators of Alzheimer’s tau pathology shared between exosomal and vesicle-free tau seeds

doi: 10.1101/2022.04.26.489622

Figure Lengend Snippet: ( a ) Schematic representation of the workflow for functional validation. Individual sgRNAs were used to silence the corresponding genes in combination with KRAB-dCas9 in tau biosensor cells (BSKRAB-KD), then treated with either sarkosyl-insoluble tau (vesicle-free tau seeds) or exosomal tau seeds for 72 hours, followed by detection and quantification of tau aggregation using FRET flow cytometry. A fraction of the same BSKRAB-KD cells was also grown for 72 hours to corroborate knockdown of protein expression using western blots. ( b-g ) Integrated FRET intensities represent levels of tau aggregation upon knocking down the different targets. The control black column (NT) is the average obtained with three independent non-targeting sgRNAs (n=3) assessed in triplicates. Control cells were compared with knockdown cells targeted individually (1, 2, and 3). Error bars represent SEM for n=3, * p< 0.05; ** p< 0.01; *** p< 0.001; **** p< 0.0001. Each single targeting sgRNA increased tau aggregation with both exosomal and vesicle-free tau seeds. ( c , d ) Interestingly, ANKLE2 and BANF1 appear to induce a stronger effect on tau aggregation induced by exosomes. ( h ) Quantitative western blot analysis of BSKRAB-KD knockdown cells. Each sgRNAs generated a protein knockdown of the targeted gene. Note that the ANKLE2-specific antibody reacts with several isoforms, including the canonical variant sized 104-117 kDa (red box outline), however, all isoforms were downregulated when the ANKLE2 locus was silenced. Similarly, the EIF1AD antibody recognized the canonical isoform of 19 kDa and one additional variant of lower molecular weight, both being silenced with the individual sgRNAs against EIF1AD. ( i ) Quantification of the extent of protein knockdown for the different targeted genes. Error bars represent SEM for n=3, *p<0.05; **p<0.01,, ****p<0.0001.

Article Snippet: The following antibodies were used: Anti-human ANKLE2 rabbit polyclonal (1:1,000; Thermo A302-965A), anti-human EIF1AD rabbit polyclonal (1:1,000; Thermo 20528-1-AP), anti-BANF1/BAF rabbit monoclonal (1:1,000; Abcam ab129184), anti-human NUSAP1 rabbit polyclonal (1:500; Abcam ab137230), anti-human VPS18 rabbit monoclonal (1:1,000; Abcam ab178689), anti-human Tau-13 mouse monoclonal (1:1,000; Covance MMS-520R), anti Tau phospho Ser422 rabbit polyclonal (1:1,000; GeneTex GTX86147), and the normalizers anti-GAPDH mouse monoclonal (1:2,000; Millipore MAB374) and anti-GAPDH rabbit polyclonal (1:2,000; Proteintech 10494-1-AP).

Techniques: Functional Assay, Biomarker Discovery, Flow Cytometry, Knockdown, Expressing, Western Blot, Control, Generated, Variant Assay, Molecular Weight

( a ) Western blot analysis of validated novel regulators using post mortem cortical brain samples from Alzheimer’s patients (AD). For the pathological data of the patients see . Antibodies against human VSP18, NUSAP1, EIF1AD and ANKLE2 were used. ( b ) Detection of human tau (Tau-13 antibody) and tau phosphorylated at Ser-422 (pS422 antibody) in the human brain samples. ( c-f ) Quantification of the protein levels in control and AD samples reveals that VPS18 ( c ), NUSAP1 ( d ) and EIF1AD ( e ) are significantly downregulated in AD patients. However, ANKLE2 levels ( f ) were not statistically different between both types of brain samples analyzed. Of note, the canonical isoform with a size of 104-117 kDa (red box outline) was the predominant isoform detected in human brains, and therefore, only this isoform was quantified. ( g ) Quantification of the ratio of pS422/hTau13 shows a substantial increase in tau phosphorylation at Ser-422 in AD samples.

Journal: bioRxiv

Article Title: Genome-wide CRISPRi screening reveals regulators of Alzheimer’s tau pathology shared between exosomal and vesicle-free tau seeds

doi: 10.1101/2022.04.26.489622

Figure Lengend Snippet: ( a ) Western blot analysis of validated novel regulators using post mortem cortical brain samples from Alzheimer’s patients (AD). For the pathological data of the patients see . Antibodies against human VSP18, NUSAP1, EIF1AD and ANKLE2 were used. ( b ) Detection of human tau (Tau-13 antibody) and tau phosphorylated at Ser-422 (pS422 antibody) in the human brain samples. ( c-f ) Quantification of the protein levels in control and AD samples reveals that VPS18 ( c ), NUSAP1 ( d ) and EIF1AD ( e ) are significantly downregulated in AD patients. However, ANKLE2 levels ( f ) were not statistically different between both types of brain samples analyzed. Of note, the canonical isoform with a size of 104-117 kDa (red box outline) was the predominant isoform detected in human brains, and therefore, only this isoform was quantified. ( g ) Quantification of the ratio of pS422/hTau13 shows a substantial increase in tau phosphorylation at Ser-422 in AD samples.

Article Snippet: The following antibodies were used: Anti-human ANKLE2 rabbit polyclonal (1:1,000; Thermo A302-965A), anti-human EIF1AD rabbit polyclonal (1:1,000; Thermo 20528-1-AP), anti-BANF1/BAF rabbit monoclonal (1:1,000; Abcam ab129184), anti-human NUSAP1 rabbit polyclonal (1:500; Abcam ab137230), anti-human VPS18 rabbit monoclonal (1:1,000; Abcam ab178689), anti-human Tau-13 mouse monoclonal (1:1,000; Covance MMS-520R), anti Tau phospho Ser422 rabbit polyclonal (1:1,000; GeneTex GTX86147), and the normalizers anti-GAPDH mouse monoclonal (1:2,000; Millipore MAB374) and anti-GAPDH rabbit polyclonal (1:2,000; Proteintech 10494-1-AP).

Techniques: Western Blot, Control, Phospho-proteomics