ChIP-seq of Sus scrofa: wild-type pEPSCs
ChIP assays were performed from 107 pEPSCs per experiment, following a previously described protocol with slight modifications (Rada-Iglesias et al., 2011). Briefly, the cells were crosslinked with 1% formaldehyde for 10 minutes at room temperature, and the reaction was quenched by adding glycine to a final concentration of 125 mM. Chromatin was then sonicated to an average size of 150C300 bp using a Bioruptor (Diagenode). A total of 1 ?g antibody was pre-incubated with magnetic beads and subsequently introduced to the sonicated chromatin. This mixture was incubated overnight at 4C. After incubation, the magnetic beads were subjected to a series of washes, and the chromatin was eluted. Crosslinks were then reversed, followed by the purification of the DNA. Library preparation was conducted utilizing the KAPA Hyper Prep Kit (KK8500) in accordance with the manufacturer's guidelines.