Pectoralis major muscle samples were collected from 35 days old chickens. Sampling (approx. 800 mg) was performed in the middle of the tissue. Samples were snap frozen in liquid nitrogen. standard Cobb500 growth condition. TM: 39.5°C and 65% RH 12h/days from embryonic days E7 to E16 and the rest of the time 37,8°C 55% RH during embryogenesis. no treatment (Control): 37,8°C and 55% RH during embryogenesis. Muscle native chromatin immunoprecipitation (ChIP) was performed as described in David et al., 2017 (doi: 10.1186/s12575-017-0059-0). Antibodies batches used: Millipore 07-449#2506493 (H3K27me3); Millipore 07-473#2717639 (H3K4me3). Sequencing libraries were prepared using the NEB Next Ultra II DNA Library Prep kit for Illumina (New England Biolabs, Ipswich, USA). Ten ng of H3K27me3 IP DNA, 10 ng of H3K4me3 IP DNA and 100 ng of input DNA were used. Agencourt AMPure XP beads (Beckman Coulter, Brea, USA) were used for the 200 bp size selection of DNA fragments. Libraries concentrations were measured by Qubit and stored at −80 °C until sequencing.
Experimental Factor: growth condition
temperature manipulation during embryogenesis
Experimental Factor: immunoprecipitate
anti-H3K27me3 (Millipore)