POGZ suppresses 2C transcriptional program and retrotransposable elements in ESCs

After centrifugation, 50 μl of supernatants were saved as input. The remainder of the chromatin solution was incubated with Dynabeads previously coupled with 10 μg ChIP grade antibodies (FLAG, Sigma, F1804; H3K4me3, Abcam, ab1012; H3K27ac, Millipore, MABE647; H3K9me3 ,Abcam, ab176916; H4K20me3, Cell Signaling Technology, 5737s; TRIM28 SETDB1) overnight at 4℃ with rotation. Next day, after 7 times washing with the wash buffer, the complexes were reverse cross-linked overnight at 65℃.

DNAs were extracted by hydroxybenzene-chloroform-isoamyl alcohol and purified by a Phase Lock Gel (Tiangen, China). For ChIP-seq, the ChIPed DNA were dissolved in 15 μl distilled water. Library constructions and deep sequencing were done by the BGI Shenzhen (Wuhan, China).

All ChIP-seq experiments were repeated at least two times. The Cleavage Under Targets and Tagmentation (CUT&Tag) experiments were performed using the 3×Flag-POGZ restoring Pogz-/- mESCs by the Yingzi and Frasergen Company (Wuhan, China) . The detailed methods have been recently described.