GSM4214115: KSHV positive Pα(+)S MSCs in MEM alpha media H3K27me3 ChIP rep3; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K27me3
Cell type
Cell type Class
Others
Cell type
Mesenchymal stem cells
NA
NA
Attributes by original data submitter
Sample
source_name
Mouse mesenchymal stem cells (MSCs) PDGFRA +
treatment
MSC media
antibody
H3K27me3
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
KSHV MEM and KSHV KS cells were crosslinked with 1% formaldehyde at room temperature for 10 min. Crosslinking was terminated by adding glycine to a final concentration of 0.125M. Cells were washed twice in ice-cold PBS and harvested by centrifugation. Every 2×107 cells were lysed in 5 ml ice-cold Farnham lysis buffer (5 mM PIPES [pH 8.0], 85 mM KCl, 0.5% NP-40) with protease inhibitors. The nuclei were spun down and resuspended in 1 ml RIPA buffer (1% NP-40, 0.5% sodium deoxycholate, and 0.3% SDS in 1×PBS) with protease inhibitors. Chromatin was sheared to about 250 bp fragments with 10 cycles using a Bioruptor Pico sonication device (Diagenode). The NEB Next Ultra II DNA Library Prep Kit for Illumina (cat. # E7645L) was used for preparing libraries with the manufacturer's protocol and associated reagents.