Polycomb Bodies Detection in Murine Fibromuscular Stroma from Skin, Skeletal Muscles, and Aortic Tissues

Methods Mol Biol. 2023:2655:125-146. doi: 10.1007/978-1-0716-3143-0_11.

Abstract

The regulation of chromatin structure depends on a dynamic, multiple mechanisms that modulate gene expression and constitute the epigenome. The Polycomb group (PcG) of proteins are epigenetic factors involved in the transcriptional repression. Among their multilevel, chromatin-associated functions, PcG proteins mediate the establishment and maintenance of higher-order structures at target genes, allowing the transmission of transcriptional programs throughout the cell cycle.In the nucleus, PcG proteins localize close to the pericentric heterochromatin forming microscopically foci, called Polycomb bodies. Here, to visualize the tissue-specific PcG distribution in the aorta, dorsal skin and hindlimb muscles, we combine a fluorescence-activated cell sorter (FACS)-based method with an immunofluorescence staining.

Keywords: Fresh tissue digestion; Immunofluorescence; Murine fibroblast characterization; Polycomb foci.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus / metabolism
  • Chromatin* / metabolism
  • Drosophila Proteins* / metabolism
  • Heterochromatin / metabolism
  • Mice
  • Muscle, Skeletal / metabolism
  • Polycomb-Group Proteins / genetics

Substances

  • Polycomb-Group Proteins
  • Chromatin
  • Heterochromatin
  • Drosophila Proteins