GSM4680907: LSEC_H3K4me3; Mus musculus; ChIP-Seq
Livers were perfused in situ via the portal vein with a 0.05% collagenase/amino acid/saccharide solution, dissected, mechanically disrupted, pooled from three mice, digested at 38 °C in collagenase/Gey's balanced salt solution and filtered through a 250 µm mesh. Non-parenchymal cells were separated by a 19.3 % Nycodenz  gradient. Afterwards, LSEC were isolated by magnetic-activated cell sorting (MACS) using anti-CD146 MicroBeads according to the manufacturers' instructions (Miltenyi Biotech). Purity of LSEC isolates was confirmed by flow cytometry and showed at least 95 % positivity for CD31 and/or Stabilin-2. Chromatin of 1 million LSEC (approx. 1 millions) was immunoprecipitated with anti-H3K4me3 (ab8580) or anti-H3K27Ac (ab4729) according to the protocol described in (PMID:31024170) After DNA purification, NGS libraries were constructed using the TruSeq ChIP library preparation kit (#IP-202-1024) according to manufacturer's recommendation. Libraries were sequenced in NextSeq550 platform using V2.5 chemistry
GEO Accession
GSM4680907