Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages
Summary: Lysine lactylation is a distinctive histone modification that plays a crucial role in epigenetic regulation and gene transcription. Here, we present a protocol for studying the genomic profile of histone lactylation with the CUT&RUN assay in tumor-associated macrophages. We describe ste...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725001728 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850202083291037696 |
|---|---|
| author | Alessandra De Leo Alessio Ugolini Filippo Veglia |
| author_facet | Alessandra De Leo Alessio Ugolini Filippo Veglia |
| author_sort | Alessandra De Leo |
| collection | DOAJ |
| description | Summary: Lysine lactylation is a distinctive histone modification that plays a crucial role in epigenetic regulation and gene transcription. Here, we present a protocol for studying the genomic profile of histone lactylation with the CUT&RUN assay in tumor-associated macrophages. We describe steps for preparing live cells, gating strategies for isolating glucose transporter type 1 + (GLUT1+) monocyte-derived macrophages (MDMs) by fluorescence-activated cell sorting (FACS), binding lactyl-specific primary antibodies, and quantifying DNA using qPCR. This protocol is applicable to both tumor-derived and in-vitro-generated bone-marrow-derived macrophages.For complete details on the use and execution of this protocol, please refer to De Leo et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-dbc8a4a679a7486f8460d71e51c0ed88 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-dbc8a4a679a7486f8460d71e51c0ed882025-08-20T02:11:50ZengElsevierSTAR Protocols2666-16672025-06-016210376610.1016/j.xpro.2025.103766Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophagesAlessandra De Leo0Alessio Ugolini1Filippo Veglia2Wistar Institute, Philadelphia, PA 19104, USA; Department of Immunology, H. Lee Moffitt Cancer Center, Tampa, FL, USA; Corresponding authorWistar Institute, Philadelphia, PA 19104, USA; Department of Immunology, H. Lee Moffitt Cancer Center, Tampa, FL, USA; Corresponding authorWistar Institute, Philadelphia, PA 19104, USA; Department of Immunology, H. Lee Moffitt Cancer Center, Tampa, FL, USA; Department of Neuro-Oncology, H. Lee Moffitt Cancer Center, Tampa, FL, USA; Corresponding authorSummary: Lysine lactylation is a distinctive histone modification that plays a crucial role in epigenetic regulation and gene transcription. Here, we present a protocol for studying the genomic profile of histone lactylation with the CUT&RUN assay in tumor-associated macrophages. We describe steps for preparing live cells, gating strategies for isolating glucose transporter type 1 + (GLUT1+) monocyte-derived macrophages (MDMs) by fluorescence-activated cell sorting (FACS), binding lactyl-specific primary antibodies, and quantifying DNA using qPCR. This protocol is applicable to both tumor-derived and in-vitro-generated bone-marrow-derived macrophages.For complete details on the use and execution of this protocol, please refer to De Leo et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725001728GenomicsImmunologyMetabolism |
| spellingShingle | Alessandra De Leo Alessio Ugolini Filippo Veglia Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages STAR Protocols Genomics Immunology Metabolism |
| title | Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages |
| title_full | Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages |
| title_fullStr | Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages |
| title_full_unstemmed | Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages |
| title_short | Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages |
| title_sort | protocol to study the genomic profile of histone lactylation with cut run assay in tumor associated macrophages |
| topic | Genomics Immunology Metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725001728 |
| work_keys_str_mv | AT alessandradeleo protocoltostudythegenomicprofileofhistonelactylationwithcutrunassayintumorassociatedmacrophages AT alessiougolini protocoltostudythegenomicprofileofhistonelactylationwithcutrunassayintumorassociatedmacrophages AT filippoveglia protocoltostudythegenomicprofileofhistonelactylationwithcutrunassayintumorassociatedmacrophages |