Control:Article Title: Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies rabbit mAb anti-LEF1 Cell Signaling Technology Cat. # 2230 rabbit mAb anti-ERG Cell Signaling Technology Cat. # 97249 chicken polyclonal anti-GFP Aves Labs Cat. # GFP-1020 rat monoclonal anti-PECAM1 BD Biosciences Cat. # 553370 goat polyclonal anti-mScarlet Biorbyt Cat. # orb1463262 rat monoclonal anti-PLVAP BD Biosciences Cat. # 553849 rabbit monoclonal anti-GLUT1 Invitrogen Cat. # MA5-31960 mouse monoclonal anti-α Smooth Muscle Actin, Alexa Fluor 647 conjugated BioLegend Cat. # 614855 rabbit monoclonal anti-GLUT1, iFluor 488 conjugated HUABIO Cat. # HA720154F rabbit monoclonal anti-ERG, Alexa Fluor 647 conjugated Abcam Cat. # ab196149 Rabbit mAb IgG XP Isotype Control Cell Signaling Technology Cat. # 66362 Alexa dye-conjugated secondary antibodies (multiple) Thermo Fisher N/A Bacterial and virus strains NEB-Stable E. coli New England BioLabs Cat# C3040H Chemicals, peptides, and recombinant proteins Cas9 protein Integrated DNA Technologies (IDT) Cat. # 1081058 NucleoBond Xtra Midi Plus EF kit TaKaRa Cat. # 740422.10 Isoflurane MP Biomedicals Cat. # ICN15195790 1 ml tuberculin syringe Becton, Dickinson Cat. # BD 309659 30 gauge x 1/2 inch PrecisionGlide Needle Becton, Dickinson Cat. # BD 305106 Trypan blue Gibco Cat. #15250061 EasyTouch Insulin Syringe (30G 1cc 5/16") Amazon N/A SeaPlaque Agarose Lonza Cat. # BMA50100 Fluoromount G EM Sciences Cat. # 17984–25 Dulbecco’s Phosphate Buffered Saline (no Ca or Mg) Gibco Cat. # 14190144 EDTA Thermo Fisher Cat. # AM9262 RNasin-Plus RNase Inhibitor Promega Cat. # N2615 Biotinylated isolectin GS-IB4 (GSL) Thermo Fisher cat. # I21414 Isolectin GS-IB4 from Griffonia simplicifolia (GSL), Alexa Fluor 488 conjugated Thermo Fisher Cat. # I21411 Ficoll PM 400 Sigma-Aldrich Cat. # F4375 Streptavidin Magnetic Beads Thermo Fisher Cat. # 88816 Spermidine Sigma Cat. # S2501 1x cOmplete, EDTA-free Protease Inhibitor Cocktail Roche Cat. # 11873580001 300 μm Nylon Mesh Filters, 47 mm diameter, Nonsterile Tisch Scientific Cat. # ME17240 20 μm filter (pluriStrainer) Cell Strainer Cat # 43-50020-03 Direct-zol RNA Microprep Kits Zymo Cat. # R2061 TRI reagent Zymo Cat. # R2050-1-50 (Continued on next page) e1 Neuron 114, 1–15.e1–e9, May 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER LunaScript RT Master Mix Kit New England BioLabs Cat. # E3025L RNase H New England BioLabs Cat. # M0297S Random 6-mer primers New England BioLabs Cat. # S1230S Novex pre-poured TBE Gels (10% acrylamide, 10 wells) Thermo Fisher Cat. # EC6275BOX GelStar Nucleic Acid Gel Stain Lonza Cat. # 50535 Spin-X Centrifuge Tube Filter CA membrane Corning Costar Cat. # 07-200-388 QIAquick Gel Extraction Kit Qiagen Cat. # 28706 Q5 High-Fidelity DNA Polymerase and Q5 High GC Enhancer New England BioLabs Cat. # M0491S CUT&RUN Assay Kit Cell Signaling Technology Cat. # 86652 DNA Purification Buffers and Spin Columns Cell Signaling Technology Cat. # 14209 SimpleChIP ChIP-seq DNA Library Prep Kit for the Illumina platform Cell Signaling Technology Cat. # 56795 AMPure XP Beads and Bead Reagents Beckman Coulter Life Sciences Cat. # A63880 Critical commercial assays DNA sequencing on a NextSeq 500 and on a NovaSeq X Plus Illumina N/A Oxford Nanopore DNA sequencing service Plasmidsaurus https://www.plasmidsaurus. com/home rAAV production Viral Tools core laboratory, Janelia Research Campus (HHMI) https://www.janelia.org/ support-team/viral-tools Deposited data CUT&RUN data (https://www.ncbi.nlm.nih. gov/sra) This paper ID PRJNA1277276 Experimental models: Organisms/strains C57BL6/J mice (pigmented) Jackson Laboratories JAX 000664 C57BL6/J mice (albino) Jackson Laboratories JAX 000058 B6J.129 R26-Cas9-GFP mice Jackson Laboratories JAX 026179 Tie2-GFP mice Jackson Laboratories JAX 008863 Oligonucleotides sgRNAs, DNA oligonucleotides, gene blocks Integrated DNA Technologies (IDT) N/A Synthetic 300-mer library Twist BioScience N/A SimpleChIP ChIP-seq Multiplex Oligonucleotides for the Illumina platform (Single Index Primers) Cell Signaling Technology Cat. # 29580 Recombinant DNA lacZ reporter plasmid Addgene, Kvon et al. 34 Cat. # 139098 pAAV-CAG-SV40NLSf-GFP-3xmiR122WPRE-HGHpA plasmid Addgene Cat. # 183775 NanoLuc Reporter Vector plasmid Promega Cat. # N1111 kiCAP-AAV-BI30 plasmid Addgene Cat # 183749 pUCmini-iCAP-PHP.B plasmid Addgene Cat. # 103002 pmU6-gRNA plasmid Addgene Cat. # 53187 pLentiCRISPRv2-blast plasmid Addgene Cat. # 98293 phH1-gRNA plasmid Addgene Cat. # 53186 ph7SK-gRNA plasmid Addgene Cat. # 53189 (Continued on next page) Neuron 114, 1–15.e1–e9, May 20, 2026 e2
Virus:Article Title: Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies rabbit mAb anti-LEF1 Cell Signaling Technology Cat. # 2230 rabbit mAb anti-ERG Cell Signaling Technology Cat. # 97249 chicken polyclonal anti-GFP Aves Labs Cat. # GFP-1020 rat monoclonal anti-PECAM1 BD Biosciences Cat. # 553370 goat polyclonal anti-mScarlet Biorbyt Cat. # orb1463262 rat monoclonal anti-PLVAP BD Biosciences Cat. # 553849 rabbit monoclonal anti-GLUT1 Invitrogen Cat. # MA5-31960 mouse monoclonal anti-α Smooth Muscle Actin, Alexa Fluor 647 conjugated BioLegend Cat. # 614855 rabbit monoclonal anti-GLUT1, iFluor 488 conjugated HUABIO Cat. # HA720154F rabbit monoclonal anti-ERG, Alexa Fluor 647 conjugated Abcam Cat. # ab196149 Rabbit mAb IgG XP Isotype Control Cell Signaling Technology Cat. # 66362 Alexa dye-conjugated secondary antibodies (multiple) Thermo Fisher N/A Bacterial and virus strains NEB-Stable E. coli New England BioLabs Cat# C3040H Chemicals, peptides, and recombinant proteins Cas9 protein Integrated DNA Technologies (IDT) Cat. # 1081058 NucleoBond Xtra Midi Plus EF kit TaKaRa Cat. # 740422.10 Isoflurane MP Biomedicals Cat. # ICN15195790 1 ml tuberculin syringe Becton, Dickinson Cat. # BD 309659 30 gauge x 1/2 inch PrecisionGlide Needle Becton, Dickinson Cat. # BD 305106 Trypan blue Gibco Cat. #15250061 EasyTouch Insulin Syringe (30G 1cc 5/16") Amazon N/A SeaPlaque Agarose Lonza Cat. # BMA50100 Fluoromount G EM Sciences Cat. # 17984–25 Dulbecco’s Phosphate Buffered Saline (no Ca or Mg) Gibco Cat. # 14190144 EDTA Thermo Fisher Cat. # AM9262 RNasin-Plus RNase Inhibitor Promega Cat. # N2615 Biotinylated isolectin GS-IB4 (GSL) Thermo Fisher cat. # I21414 Isolectin GS-IB4 from Griffonia simplicifolia (GSL), Alexa Fluor 488 conjugated Thermo Fisher Cat. # I21411 Ficoll PM 400 Sigma-Aldrich Cat. # F4375 Streptavidin Magnetic Beads Thermo Fisher Cat. # 88816 Spermidine Sigma Cat. # S2501 1x cOmplete, EDTA-free Protease Inhibitor Cocktail Roche Cat. # 11873580001 300 μm Nylon Mesh Filters, 47 mm diameter, Nonsterile Tisch Scientific Cat. # ME17240 20 μm filter (pluriStrainer) Cell Strainer Cat # 43-50020-03 Direct-zol RNA Microprep Kits Zymo Cat. # R2061 TRI reagent Zymo Cat. # R2050-1-50 (Continued on next page) e1 Neuron 114, 1–15.e1–e9, May 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER LunaScript RT Master Mix Kit New England BioLabs Cat. # E3025L RNase H New England BioLabs Cat. # M0297S Random 6-mer primers New England BioLabs Cat. # S1230S Novex pre-poured TBE Gels (10% acrylamide, 10 wells) Thermo Fisher Cat. # EC6275BOX GelStar Nucleic Acid Gel Stain Lonza Cat. # 50535 Spin-X Centrifuge Tube Filter CA membrane Corning Costar Cat. # 07-200-388 QIAquick Gel Extraction Kit Qiagen Cat. # 28706 Q5 High-Fidelity DNA Polymerase and Q5 High GC Enhancer New England BioLabs Cat. # M0491S CUT&RUN Assay Kit Cell Signaling Technology Cat. # 86652 DNA Purification Buffers and Spin Columns Cell Signaling Technology Cat. # 14209 SimpleChIP ChIP-seq DNA Library Prep Kit for the Illumina platform Cell Signaling Technology Cat. # 56795 AMPure XP Beads and Bead Reagents Beckman Coulter Life Sciences Cat. # A63880 Critical commercial assays DNA sequencing on a NextSeq 500 and on a NovaSeq X Plus Illumina N/A Oxford Nanopore DNA sequencing service Plasmidsaurus https://www.plasmidsaurus. com/home rAAV production Viral Tools core laboratory, Janelia Research Campus (HHMI) https://www.janelia.org/ support-team/viral-tools Deposited data CUT&RUN data (https://www.ncbi.nlm.nih. gov/sra) This paper ID PRJNA1277276 Experimental models: Organisms/strains C57BL6/J mice (pigmented) Jackson Laboratories JAX 000664 C57BL6/J mice (albino) Jackson Laboratories JAX 000058 B6J.129 R26-Cas9-GFP mice Jackson Laboratories JAX 026179 Tie2-GFP mice Jackson Laboratories JAX 008863 Oligonucleotides sgRNAs, DNA oligonucleotides, gene blocks Integrated DNA Technologies (IDT) N/A Synthetic 300-mer library Twist BioScience N/A SimpleChIP ChIP-seq Multiplex Oligonucleotides for the Illumina platform (Single Index Primers) Cell Signaling Technology Cat. # 29580 Recombinant DNA lacZ reporter plasmid Addgene, Kvon et al. 34 Cat. # 139098 pAAV-CAG-SV40NLSf-GFP-3xmiR122WPRE-HGHpA plasmid Addgene Cat. # 183775 NanoLuc Reporter Vector plasmid Promega Cat. # N1111 kiCAP-AAV-BI30 plasmid Addgene Cat # 183749 pUCmini-iCAP-PHP.B plasmid Addgene Cat. # 103002 pmU6-gRNA plasmid Addgene Cat. # 53187 pLentiCRISPRv2-blast plasmid Addgene Cat. # 98293 phH1-gRNA plasmid Addgene Cat. # 53186 ph7SK-gRNA plasmid Addgene Cat. # 53189 (Continued on next page) Neuron 114, 1–15.e1–e9, May 20, 2026 e2
Recombinant:Article Title: Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies rabbit mAb anti-LEF1 Cell Signaling Technology Cat. # 2230 rabbit mAb anti-ERG Cell Signaling Technology Cat. # 97249 chicken polyclonal anti-GFP Aves Labs Cat. # GFP-1020 rat monoclonal anti-PECAM1 BD Biosciences Cat. # 553370 goat polyclonal anti-mScarlet Biorbyt Cat. # orb1463262 rat monoclonal anti-PLVAP BD Biosciences Cat. # 553849 rabbit monoclonal anti-GLUT1 Invitrogen Cat. # MA5-31960 mouse monoclonal anti-α Smooth Muscle Actin, Alexa Fluor 647 conjugated BioLegend Cat. # 614855 rabbit monoclonal anti-GLUT1, iFluor 488 conjugated HUABIO Cat. # HA720154F rabbit monoclonal anti-ERG, Alexa Fluor 647 conjugated Abcam Cat. # ab196149 Rabbit mAb IgG XP Isotype Control Cell Signaling Technology Cat. # 66362 Alexa dye-conjugated secondary antibodies (multiple) Thermo Fisher N/A Bacterial and virus strains NEB-Stable E. coli New England BioLabs Cat# C3040H Chemicals, peptides, and recombinant proteins Cas9 protein Integrated DNA Technologies (IDT) Cat. # 1081058 NucleoBond Xtra Midi Plus EF kit TaKaRa Cat. # 740422.10 Isoflurane MP Biomedicals Cat. # ICN15195790 1 ml tuberculin syringe Becton, Dickinson Cat. # BD 309659 30 gauge x 1/2 inch PrecisionGlide Needle Becton, Dickinson Cat. # BD 305106 Trypan blue Gibco Cat. #15250061 EasyTouch Insulin Syringe (30G 1cc 5/16") Amazon N/A SeaPlaque Agarose Lonza Cat. # BMA50100 Fluoromount G EM Sciences Cat. # 17984–25 Dulbecco’s Phosphate Buffered Saline (no Ca or Mg) Gibco Cat. # 14190144 EDTA Thermo Fisher Cat. # AM9262 RNasin-Plus RNase Inhibitor Promega Cat. # N2615 Biotinylated isolectin GS-IB4 (GSL) Thermo Fisher cat. # I21414 Isolectin GS-IB4 from Griffonia simplicifolia (GSL), Alexa Fluor 488 conjugated Thermo Fisher Cat. # I21411 Ficoll PM 400 Sigma-Aldrich Cat. # F4375 Streptavidin Magnetic Beads Thermo Fisher Cat. # 88816 Spermidine Sigma Cat. # S2501 1x cOmplete, EDTA-free Protease Inhibitor Cocktail Roche Cat. # 11873580001 300 μm Nylon Mesh Filters, 47 mm diameter, Nonsterile Tisch Scientific Cat. # ME17240 20 μm filter (pluriStrainer) Cell Strainer Cat # 43-50020-03 Direct-zol RNA Microprep Kits Zymo Cat. # R2061 TRI reagent Zymo Cat. # R2050-1-50 (Continued on next page) e1 Neuron 114, 1–15.e1–e9, May 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER LunaScript RT Master Mix Kit New England BioLabs Cat. # E3025L RNase H New England BioLabs Cat. # M0297S Random 6-mer primers New England BioLabs Cat. # S1230S Novex pre-poured TBE Gels (10% acrylamide, 10 wells) Thermo Fisher Cat. # EC6275BOX GelStar Nucleic Acid Gel Stain Lonza Cat. # 50535 Spin-X Centrifuge Tube Filter CA membrane Corning Costar Cat. # 07-200-388 QIAquick Gel Extraction Kit Qiagen Cat. # 28706 Q5 High-Fidelity DNA Polymerase and Q5 High GC Enhancer New England BioLabs Cat. # M0491S CUT&RUN Assay Kit Cell Signaling Technology Cat. # 86652 DNA Purification Buffers and Spin Columns Cell Signaling Technology Cat. # 14209 SimpleChIP ChIP-seq DNA Library Prep Kit for the Illumina platform Cell Signaling Technology Cat. # 56795 AMPure XP Beads and Bead Reagents Beckman Coulter Life Sciences Cat. # A63880 Critical commercial assays DNA sequencing on a NextSeq 500 and on a NovaSeq X Plus Illumina N/A Oxford Nanopore DNA sequencing service Plasmidsaurus https://www.plasmidsaurus. com/home rAAV production Viral Tools core laboratory, Janelia Research Campus (HHMI) https://www.janelia.org/ support-team/viral-tools Deposited data CUT&RUN data (https://www.ncbi.nlm.nih. gov/sra) This paper ID PRJNA1277276 Experimental models: Organisms/strains C57BL6/J mice (pigmented) Jackson Laboratories JAX 000664 C57BL6/J mice (albino) Jackson Laboratories JAX 000058 B6J.129 R26-Cas9-GFP mice Jackson Laboratories JAX 026179 Tie2-GFP mice Jackson Laboratories JAX 008863 Oligonucleotides sgRNAs, DNA oligonucleotides, gene blocks Integrated DNA Technologies (IDT) N/A Synthetic 300-mer library Twist BioScience N/A SimpleChIP ChIP-seq Multiplex Oligonucleotides for the Illumina platform (Single Index Primers) Cell Signaling Technology Cat. # 29580 Recombinant DNA lacZ reporter plasmid Addgene, Kvon et al. 34 Cat. # 139098 pAAV-CAG-SV40NLSf-GFP-3xmiR122WPRE-HGHpA plasmid Addgene Cat. # 183775 NanoLuc Reporter Vector plasmid Promega Cat. # N1111 kiCAP-AAV-BI30 plasmid Addgene Cat # 183749 pUCmini-iCAP-PHP.B plasmid Addgene Cat. # 103002 pmU6-gRNA plasmid Addgene Cat. # 53187 pLentiCRISPRv2-blast plasmid Addgene Cat. # 98293 phH1-gRNA plasmid Addgene Cat. # 53186 ph7SK-gRNA plasmid Addgene Cat. # 53189 (Continued on next page) Neuron 114, 1–15.e1–e9, May 20, 2026 e2
Saline:Article Title: Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies rabbit mAb anti-LEF1 Cell Signaling Technology Cat. # 2230 rabbit mAb anti-ERG Cell Signaling Technology Cat. # 97249 chicken polyclonal anti-GFP Aves Labs Cat. # GFP-1020 rat monoclonal anti-PECAM1 BD Biosciences Cat. # 553370 goat polyclonal anti-mScarlet Biorbyt Cat. # orb1463262 rat monoclonal anti-PLVAP BD Biosciences Cat. # 553849 rabbit monoclonal anti-GLUT1 Invitrogen Cat. # MA5-31960 mouse monoclonal anti-α Smooth Muscle Actin, Alexa Fluor 647 conjugated BioLegend Cat. # 614855 rabbit monoclonal anti-GLUT1, iFluor 488 conjugated HUABIO Cat. # HA720154F rabbit monoclonal anti-ERG, Alexa Fluor 647 conjugated Abcam Cat. # ab196149 Rabbit mAb IgG XP Isotype Control Cell Signaling Technology Cat. # 66362 Alexa dye-conjugated secondary antibodies (multiple) Thermo Fisher N/A Bacterial and virus strains NEB-Stable E. coli New England BioLabs Cat# C3040H Chemicals, peptides, and recombinant proteins Cas9 protein Integrated DNA Technologies (IDT) Cat. # 1081058 NucleoBond Xtra Midi Plus EF kit TaKaRa Cat. # 740422.10 Isoflurane MP Biomedicals Cat. # ICN15195790 1 ml tuberculin syringe Becton, Dickinson Cat. # BD 309659 30 gauge x 1/2 inch PrecisionGlide Needle Becton, Dickinson Cat. # BD 305106 Trypan blue Gibco Cat. #15250061 EasyTouch Insulin Syringe (30G 1cc 5/16") Amazon N/A SeaPlaque Agarose Lonza Cat. # BMA50100 Fluoromount G EM Sciences Cat. # 17984–25 Dulbecco’s Phosphate Buffered Saline (no Ca or Mg) Gibco Cat. # 14190144 EDTA Thermo Fisher Cat. # AM9262 RNasin-Plus RNase Inhibitor Promega Cat. # N2615 Biotinylated isolectin GS-IB4 (GSL) Thermo Fisher cat. # I21414 Isolectin GS-IB4 from Griffonia simplicifolia (GSL), Alexa Fluor 488 conjugated Thermo Fisher Cat. # I21411 Ficoll PM 400 Sigma-Aldrich Cat. # F4375 Streptavidin Magnetic Beads Thermo Fisher Cat. # 88816 Spermidine Sigma Cat. # S2501 1x cOmplete, EDTA-free Protease Inhibitor Cocktail Roche Cat. # 11873580001 300 μm Nylon Mesh Filters, 47 mm diameter, Nonsterile Tisch Scientific Cat. # ME17240 20 μm filter (pluriStrainer) Cell Strainer Cat # 43-50020-03 Direct-zol RNA Microprep Kits Zymo Cat. # R2061 TRI reagent Zymo Cat. # R2050-1-50 (Continued on next page) e1 Neuron 114, 1–15.e1–e9, May 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER LunaScript RT Master Mix Kit New England BioLabs Cat. # E3025L RNase H New England BioLabs Cat. # M0297S Random 6-mer primers New England BioLabs Cat. # S1230S Novex pre-poured TBE Gels (10% acrylamide, 10 wells) Thermo Fisher Cat. # EC6275BOX GelStar Nucleic Acid Gel Stain Lonza Cat. # 50535 Spin-X Centrifuge Tube Filter CA membrane Corning Costar Cat. # 07-200-388 QIAquick Gel Extraction Kit Qiagen Cat. # 28706 Q5 High-Fidelity DNA Polymerase and Q5 High GC Enhancer New England BioLabs Cat. # M0491S CUT&RUN Assay Kit Cell Signaling Technology Cat. # 86652 DNA Purification Buffers and Spin Columns Cell Signaling Technology Cat. # 14209 SimpleChIP ChIP-seq DNA Library Prep Kit for the Illumina platform Cell Signaling Technology Cat. # 56795 AMPure XP Beads and Bead Reagents Beckman Coulter Life Sciences Cat. # A63880 Critical commercial assays DNA sequencing on a NextSeq 500 and on a NovaSeq X Plus Illumina N/A Oxford Nanopore DNA sequencing service Plasmidsaurus https://www.plasmidsaurus. com/home rAAV production Viral Tools core laboratory, Janelia Research Campus (HHMI) https://www.janelia.org/ support-team/viral-tools Deposited data CUT&RUN data (https://www.ncbi.nlm.nih. gov/sra) This paper ID PRJNA1277276 Experimental models: Organisms/strains C57BL6/J mice (pigmented) Jackson Laboratories JAX 000664 C57BL6/J mice (albino) Jackson Laboratories JAX 000058 B6J.129 R26-Cas9-GFP mice Jackson Laboratories JAX 026179 Tie2-GFP mice Jackson Laboratories JAX 008863 Oligonucleotides sgRNAs, DNA oligonucleotides, gene blocks Integrated DNA Technologies (IDT) N/A Synthetic 300-mer library Twist BioScience N/A SimpleChIP ChIP-seq Multiplex Oligonucleotides for the Illumina platform (Single Index Primers) Cell Signaling Technology Cat. # 29580 Recombinant DNA lacZ reporter plasmid Addgene, Kvon et al. 34 Cat. # 139098 pAAV-CAG-SV40NLSf-GFP-3xmiR122WPRE-HGHpA plasmid Addgene Cat. # 183775 NanoLuc Reporter Vector plasmid Promega Cat. # N1111 kiCAP-AAV-BI30 plasmid Addgene Cat # 183749 pUCmini-iCAP-PHP.B plasmid Addgene Cat. # 103002 pmU6-gRNA plasmid Addgene Cat. # 53187 pLentiCRISPRv2-blast plasmid Addgene Cat. # 98293 phH1-gRNA plasmid Addgene Cat. # 53186 ph7SK-gRNA plasmid Addgene Cat. # 53189 (Continued on next page) Neuron 114, 1–15.e1–e9, May 20, 2026 e2
Magnetic Beads:Article Title: Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies rabbit mAb anti-LEF1 Cell Signaling Technology Cat. # 2230 rabbit mAb anti-ERG Cell Signaling Technology Cat. # 97249 chicken polyclonal anti-GFP Aves Labs Cat. # GFP-1020 rat monoclonal anti-PECAM1 BD Biosciences Cat. # 553370 goat polyclonal anti-mScarlet Biorbyt Cat. # orb1463262 rat monoclonal anti-PLVAP BD Biosciences Cat. # 553849 rabbit monoclonal anti-GLUT1 Invitrogen Cat. # MA5-31960 mouse monoclonal anti-α Smooth Muscle Actin, Alexa Fluor 647 conjugated BioLegend Cat. # 614855 rabbit monoclonal anti-GLUT1, iFluor 488 conjugated HUABIO Cat. # HA720154F rabbit monoclonal anti-ERG, Alexa Fluor 647 conjugated Abcam Cat. # ab196149 Rabbit mAb IgG XP Isotype Control Cell Signaling Technology Cat. # 66362 Alexa dye-conjugated secondary antibodies (multiple) Thermo Fisher N/A Bacterial and virus strains NEB-Stable E. coli New England BioLabs Cat# C3040H Chemicals, peptides, and recombinant proteins Cas9 protein Integrated DNA Technologies (IDT) Cat. # 1081058 NucleoBond Xtra Midi Plus EF kit TaKaRa Cat. # 740422.10 Isoflurane MP Biomedicals Cat. # ICN15195790 1 ml tuberculin syringe Becton, Dickinson Cat. # BD 309659 30 gauge x 1/2 inch PrecisionGlide Needle Becton, Dickinson Cat. # BD 305106 Trypan blue Gibco Cat. #15250061 EasyTouch Insulin Syringe (30G 1cc 5/16") Amazon N/A SeaPlaque Agarose Lonza Cat. # BMA50100 Fluoromount G EM Sciences Cat. # 17984–25 Dulbecco’s Phosphate Buffered Saline (no Ca or Mg) Gibco Cat. # 14190144 EDTA Thermo Fisher Cat. # AM9262 RNasin-Plus RNase Inhibitor Promega Cat. # N2615 Biotinylated isolectin GS-IB4 (GSL) Thermo Fisher cat. # I21414 Isolectin GS-IB4 from Griffonia simplicifolia (GSL), Alexa Fluor 488 conjugated Thermo Fisher Cat. # I21411 Ficoll PM 400 Sigma-Aldrich Cat. # F4375 Streptavidin Magnetic Beads Thermo Fisher Cat. # 88816 Spermidine Sigma Cat. # S2501 1x cOmplete, EDTA-free Protease Inhibitor Cocktail Roche Cat. # 11873580001 300 μm Nylon Mesh Filters, 47 mm diameter, Nonsterile Tisch Scientific Cat. # ME17240 20 μm filter (pluriStrainer) Cell Strainer Cat # 43-50020-03 Direct-zol RNA Microprep Kits Zymo Cat. # R2061 TRI reagent Zymo Cat. # R2050-1-50 (Continued on next page) e1 Neuron 114, 1–15.e1–e9, May 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER LunaScript RT Master Mix Kit New England BioLabs Cat. # E3025L RNase H New England BioLabs Cat. # M0297S Random 6-mer primers New England BioLabs Cat. # S1230S Novex pre-poured TBE Gels (10% acrylamide, 10 wells) Thermo Fisher Cat. # EC6275BOX GelStar Nucleic Acid Gel Stain Lonza Cat. # 50535 Spin-X Centrifuge Tube Filter CA membrane Corning Costar Cat. # 07-200-388 QIAquick Gel Extraction Kit Qiagen Cat. # 28706 Q5 High-Fidelity DNA Polymerase and Q5 High GC Enhancer New England BioLabs Cat. # M0491S CUT&RUN Assay Kit Cell Signaling Technology Cat. # 86652 DNA Purification Buffers and Spin Columns Cell Signaling Technology Cat. # 14209 SimpleChIP ChIP-seq DNA Library Prep Kit for the Illumina platform Cell Signaling Technology Cat. # 56795 AMPure XP Beads and Bead Reagents Beckman Coulter Life Sciences Cat. # A63880 Critical commercial assays DNA sequencing on a NextSeq 500 and on a NovaSeq X Plus Illumina N/A Oxford Nanopore DNA sequencing service Plasmidsaurus https://www.plasmidsaurus. com/home rAAV production Viral Tools core laboratory, Janelia Research Campus (HHMI) https://www.janelia.org/ support-team/viral-tools Deposited data CUT&RUN data (https://www.ncbi.nlm.nih. gov/sra) This paper ID PRJNA1277276 Experimental models: Organisms/strains C57BL6/J mice (pigmented) Jackson Laboratories JAX 000664 C57BL6/J mice (albino) Jackson Laboratories JAX 000058 B6J.129 R26-Cas9-GFP mice Jackson Laboratories JAX 026179 Tie2-GFP mice Jackson Laboratories JAX 008863 Oligonucleotides sgRNAs, DNA oligonucleotides, gene blocks Integrated DNA Technologies (IDT) N/A Synthetic 300-mer library Twist BioScience N/A SimpleChIP ChIP-seq Multiplex Oligonucleotides for the Illumina platform (Single Index Primers) Cell Signaling Technology Cat. # 29580 Recombinant DNA lacZ reporter plasmid Addgene, Kvon et al. 34 Cat. # 139098 pAAV-CAG-SV40NLSf-GFP-3xmiR122WPRE-HGHpA plasmid Addgene Cat. # 183775 NanoLuc Reporter Vector plasmid Promega Cat. # N1111 kiCAP-AAV-BI30 plasmid Addgene Cat # 183749 pUCmini-iCAP-PHP.B plasmid Addgene Cat. # 103002 pmU6-gRNA plasmid Addgene Cat. # 53187 pLentiCRISPRv2-blast plasmid Addgene Cat. # 98293 phH1-gRNA plasmid Addgene Cat. # 53186 ph7SK-gRNA plasmid Addgene Cat. # 53189 (Continued on next page) Neuron 114, 1–15.e1–e9, May 20, 2026 e2
Protease Inhibitor:Article Title: Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies rabbit mAb anti-LEF1 Cell Signaling Technology Cat. # 2230 rabbit mAb anti-ERG Cell Signaling Technology Cat. # 97249 chicken polyclonal anti-GFP Aves Labs Cat. # GFP-1020 rat monoclonal anti-PECAM1 BD Biosciences Cat. # 553370 goat polyclonal anti-mScarlet Biorbyt Cat. # orb1463262 rat monoclonal anti-PLVAP BD Biosciences Cat. # 553849 rabbit monoclonal anti-GLUT1 Invitrogen Cat. # MA5-31960 mouse monoclonal anti-α Smooth Muscle Actin, Alexa Fluor 647 conjugated BioLegend Cat. # 614855 rabbit monoclonal anti-GLUT1, iFluor 488 conjugated HUABIO Cat. # HA720154F rabbit monoclonal anti-ERG, Alexa Fluor 647 conjugated Abcam Cat. # ab196149 Rabbit mAb IgG XP Isotype Control Cell Signaling Technology Cat. # 66362 Alexa dye-conjugated secondary antibodies (multiple) Thermo Fisher N/A Bacterial and virus strains NEB-Stable E. coli New England BioLabs Cat# C3040H Chemicals, peptides, and recombinant proteins Cas9 protein Integrated DNA Technologies (IDT) Cat. # 1081058 NucleoBond Xtra Midi Plus EF kit TaKaRa Cat. # 740422.10 Isoflurane MP Biomedicals Cat. # ICN15195790 1 ml tuberculin syringe Becton, Dickinson Cat. # BD 309659 30 gauge x 1/2 inch PrecisionGlide Needle Becton, Dickinson Cat. # BD 305106 Trypan blue Gibco Cat. #15250061 EasyTouch Insulin Syringe (30G 1cc 5/16") Amazon N/A SeaPlaque Agarose Lonza Cat. # BMA50100 Fluoromount G EM Sciences Cat. # 17984–25 Dulbecco’s Phosphate Buffered Saline (no Ca or Mg) Gibco Cat. # 14190144 EDTA Thermo Fisher Cat. # AM9262 RNasin-Plus RNase Inhibitor Promega Cat. # N2615 Biotinylated isolectin GS-IB4 (GSL) Thermo Fisher cat. # I21414 Isolectin GS-IB4 from Griffonia simplicifolia (GSL), Alexa Fluor 488 conjugated Thermo Fisher Cat. # I21411 Ficoll PM 400 Sigma-Aldrich Cat. # F4375 Streptavidin Magnetic Beads Thermo Fisher Cat. # 88816 Spermidine Sigma Cat. # S2501 1x cOmplete, EDTA-free Protease Inhibitor Cocktail Roche Cat. # 11873580001 300 μm Nylon Mesh Filters, 47 mm diameter, Nonsterile Tisch Scientific Cat. # ME17240 20 μm filter (pluriStrainer) Cell Strainer Cat # 43-50020-03 Direct-zol RNA Microprep Kits Zymo Cat. # R2061 TRI reagent Zymo Cat. # R2050-1-50 (Continued on next page) e1 Neuron 114, 1–15.e1–e9, May 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER LunaScript RT Master Mix Kit New England BioLabs Cat. # E3025L RNase H New England BioLabs Cat. # M0297S Random 6-mer primers New England BioLabs Cat. # S1230S Novex pre-poured TBE Gels (10% acrylamide, 10 wells) Thermo Fisher Cat. # EC6275BOX GelStar Nucleic Acid Gel Stain Lonza Cat. # 50535 Spin-X Centrifuge Tube Filter CA membrane Corning Costar Cat. # 07-200-388 QIAquick Gel Extraction Kit Qiagen Cat. # 28706 Q5 High-Fidelity DNA Polymerase and Q5 High GC Enhancer New England BioLabs Cat. # M0491S CUT&RUN Assay Kit Cell Signaling Technology Cat. # 86652 DNA Purification Buffers and Spin Columns Cell Signaling Technology Cat. # 14209 SimpleChIP ChIP-seq DNA Library Prep Kit for the Illumina platform Cell Signaling Technology Cat. # 56795 AMPure XP Beads and Bead Reagents Beckman Coulter Life Sciences Cat. # A63880 Critical commercial assays DNA sequencing on a NextSeq 500 and on a NovaSeq X Plus Illumina N/A Oxford Nanopore DNA sequencing service Plasmidsaurus https://www.plasmidsaurus. com/home rAAV production Viral Tools core laboratory, Janelia Research Campus (HHMI) https://www.janelia.org/ support-team/viral-tools Deposited data CUT&RUN data (https://www.ncbi.nlm.nih. gov/sra) This paper ID PRJNA1277276 Experimental models: Organisms/strains C57BL6/J mice (pigmented) Jackson Laboratories JAX 000664 C57BL6/J mice (albino) Jackson Laboratories JAX 000058 B6J.129 R26-Cas9-GFP mice Jackson Laboratories JAX 026179 Tie2-GFP mice Jackson Laboratories JAX 008863 Oligonucleotides sgRNAs, DNA oligonucleotides, gene blocks Integrated DNA Technologies (IDT) N/A Synthetic 300-mer library Twist BioScience N/A SimpleChIP ChIP-seq Multiplex Oligonucleotides for the Illumina platform (Single Index Primers) Cell Signaling Technology Cat. # 29580 Recombinant DNA lacZ reporter plasmid Addgene, Kvon et al. 34 Cat. # 139098 pAAV-CAG-SV40NLSf-GFP-3xmiR122WPRE-HGHpA plasmid Addgene Cat. # 183775 NanoLuc Reporter Vector plasmid Promega Cat. # N1111 kiCAP-AAV-BI30 plasmid Addgene Cat # 183749 pUCmini-iCAP-PHP.B plasmid Addgene Cat. # 103002 pmU6-gRNA plasmid Addgene Cat. # 53187 pLentiCRISPRv2-blast plasmid Addgene Cat. # 98293 phH1-gRNA plasmid Addgene Cat. # 53186 ph7SK-gRNA plasmid Addgene Cat. # 53189 (Continued on next page) Neuron 114, 1–15.e1–e9, May 20, 2026 e2
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