biorad gene pulse Search Results


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biorad gene pulse electroporation  (Bio-Rad)
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Bio-Rad biorad gene pulse electroporation
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genepulser xcell  (Bio-Rad)
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chef mappertm xa pulsed field electrophoresis system  (Bio-Rad)
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pulsed field electrophoresis gel  (Bio-Rad)
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Bio-Rad pulsed field electrophoresis gel
Pulsed Field Electrophoresis Gel, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti h3 antibody  (Bio-Rad)
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Bio-Rad anti h3 antibody
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
Anti H3 Antibody, supplied by Bio-Rad, 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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x cell gene pulse electroporator  (Bio-Rad)
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Bio-Rad x cell gene pulse electroporator
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
X Cell Gene Pulse Electroporator, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cells by electroporation  (Bio-Rad)
93
Bio-Rad cells by electroporation
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
Cells By Electroporation, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gene pulsertm apparatus  (Bio-Rad)
96
Bio-Rad gene pulsertm apparatus
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
Gene Pulsertm Apparatus, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gene pulser xcelltm eukaryotic system instrument  (Bio-Rad)
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Bio-Rad gene pulser xcelltm eukaryotic system instrument
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
Gene Pulser Xcelltm Eukaryotic System Instrument, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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biorad genepulserxcell  (Bio-Rad)
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Bio-Rad biorad genepulserxcell
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
Biorad Genepulserxcell, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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kv pulse  (Bio-Rad)
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Bio-Rad kv pulse
Fig. 1 | TG2 is a writer, eraser and exchanger of <t>H3</t> monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors
Kv Pulse, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Fig. 1 | TG2 is a writer, eraser and exchanger of H3 monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors

Journal: Nature

Article Title: Bidirectional histone monoaminylation dynamics regulate neural rhythmicity.

doi: 10.1038/s41586-024-08371-3

Figure Lengend Snippet: Fig. 1 | TG2 is a writer, eraser and exchanger of H3 monoaminylations. a, TG2-mediated covalent modification of H3Q5 by 5-PT, and its visualization by CuAAC-meditated Cy5 conjugation. b, 5-PT pulse–chase in HeLa cells treated with or without TG2 inhibitors (ERW1041E or ZDON) after removal of 5-PT. Western blotting was performed for Cy5 (that is, H3 serotonylation) and TG2. c, 5-PT pulse–chase experiment in HEK293T cells transfected with WT TG2 and treated with or without TG2 inhibitors after removal of 5-PT. Western blotting was performed for Cy5 and TG2. d, Suggested mechanism for TG2-mediated H3 monoaminylation writing, erasing and exchange. e, LC–MS analysis of modified H3 peptides (as well as a deamidated H3Q5E peptide standard) (xi) after incubation with cellular lysates expressing WT TG2 versus TG2(C277A) (i–iv) or recombinant WT TG2 versus TG2(C277A) (v–x). H3Q5his peptide was incubated with WT TG2 in the presence or absence of replacement monoamine donors (xii (serotonin) versus xiii (dopamine)), demonstrating WT TG2-mediated deamidation of H3 monoaminylations in the absence of replacement donors

Article Snippet: Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Primary antibodies used in this study: Rabbit Anti-TGM2 1:500 (brain) Abcam CUB 7402 Rabbit Anti-GAPDH 1:1000 (Western) Thermo-Fisher PA1-16777 1: 1000 (in vitro and in cellulo) CST (3557S) Chicken Anti-H3 1: 1000 Abcam (ab134198) Mouse Anti-H3 1: 1000 Abcam (ab10799) Mouse Anti-Actin 1: 1000 CST (3700S) Rabbit Anti-H3Q5ser 1: 1000 Millipore (ABE1791) Rabbit Anti-H3Q5dop 1: 1000 Millipore (ABE2588) Rabbit Anti-H3Q5his 1: 200 (brain) 1:1000 (in vitro and in cellulo) Millipore (ABE2578) Rabbit Anti-H3K4me3Q5ser 1:50 (CUT&RUN), 1:500 (Western) Millipore (ABE2605) Rabbit Anti-H3K4me3Q5his 1:50 (CUT&RUN), 1:500 (Western) Millipore (ABE2570) Rabbit Anti-H3K4me3 1:1000 Abcam (ab8580) 4 nature portfolio | reporting sum m ary M arch 2021 Rabbit Anti-H3K4me3 1:50 (CUT&RUN), 1:500 (Western) Epicypher (13-0041) Rabbit Anti-H3K4me2 1:1000 Abcam (ab7766) Rabbit Anti-H3K4me2 1:50 (CUT&RUN) Active Motif (39141) Rabbit Anti-H3K27ac 1:1000 Active Motif (AB_2614979) Rabbit Anti-H3 1:50000 (brain) Abcam (ab1791) Rabbit Anti-WDR5 1:50 (CUT&RUN) CST (13105) Rabbit Anti-WDR5 1:1000 Abcam (ab307664) Mouse Anti-FLAG M2 1:1000 Millipore Sigma (F1804) Chicken Anti-6XHIS 1:1000 Invitrogen (PA19531) Rabbit Anti-HDC 1:1000 ARP (03–16045) Rabbit Anti-NeuN 1:1000 Millipore Sigma (MAB377) Donkey Anti-Chicken IRDye 800CW 1: 15000 (Li-Cor 926-32218) Goat anti-Chicken IgY (H+L) Secondary Antibody, Alexa FluorTM 488, 1:500 Invitrogen (for H3 peptide quantification) (Thermo Fisher A-11039) Goat Anti-Mouse IRDye 680RD 1: 15000 (Li-Cor 926-68070) Goat Anti-Mouse IRDye 800CW 1: 15000 (Li-Cor 926-32210) Goat Anti-Rabbit IRDye 800CW 1: 15000 (Li-Cor 926-32211) Goat Anti-Rabbit IRDye 680RD 1: 15000 (Li-Cor 926-68071) Goat Anti-Rabbit Horseradish Peroxidase 1:10000 1:50000 (for anti-H3 antibody) (BioRad 1706515) Sheep Anti-Mouse Horseradish Peroxidase 1:5000 (Cytiva RPN4201) Donkey Anti-Rabbit Horseradish Peroxidase 1:5000 ( Cytiva NA934) Validation All antibodies used in this study (all of which have been commercially validated (see manufacturers website) or described in previous publications) were validated in cells/tissues via immunoblotting, IPs or ICC/IHC/IF prior to experimentation.

Techniques: Modification, Conjugation Assay, Pulse Chase, Western Blot, Transfection, Liquid Chromatography with Mass Spectroscopy, Incubation, Expressing, Recombinant

Fig. 3 | Neural H3Q5 monoaminylations are diurnally rhythmic. a, RNA-seq analysis of the mouse TMN across ZT (JTKcycle, Padj < 0.05). n = 3 biological replicates per timepoint. b, Ontology analysis (P < 0.05) of RNA-seq data from a. c, Known circadian genes identified in a. d, CUT&RUN–seq enrichment for H3K4me3Q5his across ZT at genic loci in TMN. n = 3 biological replicates per timepoint (JTKcycle, Padj < 0.05). e, Ontology analysis (P < 0.05) comparing enrichment of CLOCK-mediated gene targets for genes displaying rhythmic H3K4me3Q5his only versus circadian genes not displaying rhythmic H3K4me3Q5his and/or gene expression. f, The average signal intensity ±500 bp from the transcription start site (TSS) across ZT for H3 monoaminylations and WDR5. g, The average signal intensity ±500 bp from the TSS across ZT for H3K4me3Q5his and H3K4me3. h, IGV tracks for Per2 across ZT for H3 monoaminylations versus WDR5 versus H3K4me2/3. i, Overlap of rhythmic H3 monoaminylation genes with WDR5-enriched genes at ZT16 (left), and ontology

Journal: Nature

Article Title: Bidirectional histone monoaminylation dynamics regulate neural rhythmicity.

doi: 10.1038/s41586-024-08371-3

Figure Lengend Snippet: Fig. 3 | Neural H3Q5 monoaminylations are diurnally rhythmic. a, RNA-seq analysis of the mouse TMN across ZT (JTKcycle, Padj < 0.05). n = 3 biological replicates per timepoint. b, Ontology analysis (P < 0.05) of RNA-seq data from a. c, Known circadian genes identified in a. d, CUT&RUN–seq enrichment for H3K4me3Q5his across ZT at genic loci in TMN. n = 3 biological replicates per timepoint (JTKcycle, Padj < 0.05). e, Ontology analysis (P < 0.05) comparing enrichment of CLOCK-mediated gene targets for genes displaying rhythmic H3K4me3Q5his only versus circadian genes not displaying rhythmic H3K4me3Q5his and/or gene expression. f, The average signal intensity ±500 bp from the transcription start site (TSS) across ZT for H3 monoaminylations and WDR5. g, The average signal intensity ±500 bp from the TSS across ZT for H3K4me3Q5his and H3K4me3. h, IGV tracks for Per2 across ZT for H3 monoaminylations versus WDR5 versus H3K4me2/3. i, Overlap of rhythmic H3 monoaminylation genes with WDR5-enriched genes at ZT16 (left), and ontology

Article Snippet: Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Primary antibodies used in this study: Rabbit Anti-TGM2 1:500 (brain) Abcam CUB 7402 Rabbit Anti-GAPDH 1:1000 (Western) Thermo-Fisher PA1-16777 1: 1000 (in vitro and in cellulo) CST (3557S) Chicken Anti-H3 1: 1000 Abcam (ab134198) Mouse Anti-H3 1: 1000 Abcam (ab10799) Mouse Anti-Actin 1: 1000 CST (3700S) Rabbit Anti-H3Q5ser 1: 1000 Millipore (ABE1791) Rabbit Anti-H3Q5dop 1: 1000 Millipore (ABE2588) Rabbit Anti-H3Q5his 1: 200 (brain) 1:1000 (in vitro and in cellulo) Millipore (ABE2578) Rabbit Anti-H3K4me3Q5ser 1:50 (CUT&RUN), 1:500 (Western) Millipore (ABE2605) Rabbit Anti-H3K4me3Q5his 1:50 (CUT&RUN), 1:500 (Western) Millipore (ABE2570) Rabbit Anti-H3K4me3 1:1000 Abcam (ab8580) 4 nature portfolio | reporting sum m ary M arch 2021 Rabbit Anti-H3K4me3 1:50 (CUT&RUN), 1:500 (Western) Epicypher (13-0041) Rabbit Anti-H3K4me2 1:1000 Abcam (ab7766) Rabbit Anti-H3K4me2 1:50 (CUT&RUN) Active Motif (39141) Rabbit Anti-H3K27ac 1:1000 Active Motif (AB_2614979) Rabbit Anti-H3 1:50000 (brain) Abcam (ab1791) Rabbit Anti-WDR5 1:50 (CUT&RUN) CST (13105) Rabbit Anti-WDR5 1:1000 Abcam (ab307664) Mouse Anti-FLAG M2 1:1000 Millipore Sigma (F1804) Chicken Anti-6XHIS 1:1000 Invitrogen (PA19531) Rabbit Anti-HDC 1:1000 ARP (03–16045) Rabbit Anti-NeuN 1:1000 Millipore Sigma (MAB377) Donkey Anti-Chicken IRDye 800CW 1: 15000 (Li-Cor 926-32218) Goat anti-Chicken IgY (H+L) Secondary Antibody, Alexa FluorTM 488, 1:500 Invitrogen (for H3 peptide quantification) (Thermo Fisher A-11039) Goat Anti-Mouse IRDye 680RD 1: 15000 (Li-Cor 926-68070) Goat Anti-Mouse IRDye 800CW 1: 15000 (Li-Cor 926-32210) Goat Anti-Rabbit IRDye 800CW 1: 15000 (Li-Cor 926-32211) Goat Anti-Rabbit IRDye 680RD 1: 15000 (Li-Cor 926-68071) Goat Anti-Rabbit Horseradish Peroxidase 1:10000 1:50000 (for anti-H3 antibody) (BioRad 1706515) Sheep Anti-Mouse Horseradish Peroxidase 1:5000 (Cytiva RPN4201) Donkey Anti-Rabbit Horseradish Peroxidase 1:5000 ( Cytiva NA934) Validation All antibodies used in this study (all of which have been commercially validated (see manufacturers website) or described in previous publications) were validated in cells/tissues via immunoblotting, IPs or ICC/IHC/IF prior to experimentation.

Techniques: RNA Sequencing, Gene Expression