GSM2113332: RNAPII 06hpi campothecin; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Unclassified
Antigen
Unclassified
Cell type
Cell type Class
Lung
Cell type
A549
Primary Tissue
Lung
Tissue Diagnosis
Carcinoma
Attributes by original data submitter
Sample
source_name
adenocarcinomic human alveolar basal epithelial cells
cell type
A549
viral strain
A/PR/8/34 deltaNS1
treatment
campothecin
timepoint
06 hours post-infection
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Crosslinking was performed for 10 minutes. For sonication, we used a refrigerated Bioruptor (Diagenode), which we optimized to generate DNA fragments of approximately 200– 1,000 base pair (bp). Lysates were pre-cleared for 3 hours using the appropriate isotype-matched control antibody (rabbit IgG; Cell Signaling) or anti-mouse IgG (Cell Signaling). The anti-RNA polymerase II (RNAPII) (clone 8WG16; Covance/BioLegend) antibodies were coupled with magnetic paramagnetic beads (Dynabeads® M-280 Sheep anti- 21 Mouse IgG; ThermoFisher Scientific) bound to anti–mouse IgG or anti–rabbit IgG for 6 hours. Antibody-bound beads and chromatin were then immune-precipitated overnight at 4 °C with rotation. After washing, reverse crosslinking was carried out overnight at 65°C. After digestion with RNase and proteinase K (Roche), DNA was isolated with a MinElute kit (Qiagen) and used for downstream applications.