LPS-stimulated THP1 monocytic cell line
Monocytic THP-1 cells were cultured in growth media (RPMI supplemented with 10% (v:v) FCS, 2 mM L-glutamine, 100 U/ml penicillin, 100 µg/ml streptomycin and 50 nM of 2-Mercaptoethanol (all GIBCO, Life Technologies) and incubated in a 37ºC, 5% (v:v) CO2 humidified incubator. Cell were stimulated for 4h with/without 1 µg/ml LPS (E. coli 055:B5, Sigma Aldrich). Total RNA was extracted using the RNeasy kit (Qiagen), which included an on-column DNase treatment (Qiagen), according to the manufacturer's guideline. RNA concentration was determined using the Qubit 2.0 (Life Technologies). RNA quality was measured using the Agilent Bioanalyser and produced RIN values of >8.0. Sample were dispatched to BGI Genomics (Hong Kong, China) where they underwent Illumina TruSeq polyA library preparation prior to 100 nucleotide paired-end, stranded sequencing.