GSM7070493: Cntrl_198_LPS; Homo sapiens; RNA-Seq
500.000 cells of the monocyte-enriched cell suspension were used for RNA extraction (250.000 for medium stimulated, 250.000 for LPS stimulated). The cells were spun down at 500g for 5 minutes. Supernatant was removed and the cell pellet was resuspended with 500µL TrIzoL solution. The resuspended solution was stored at -80C until RNA isolation. RNA was isolated using RNeasy Micro Kit (Qiagen) with adaptation of the first step for TriZOL samples, according to manufacturer's protocol, and included on column DNAse treatment. RNA quality was assessed on the Agilent 2100 analyzer. cDNA library construction and sequencing were performed at Beijing Genomics Institute using the BGISEQ-500 platform with a starting input of 200ng. Libraries were sequenced on the BGISeq-500 platform generating an average of 20 million reads per sample. To infer gene expression levels, RNA-seq reads were aligned to hg19 human transcriptome using Bowtie. Quantification of gene expression levels as RPKM was performed using MMSEQ. Reads/transcripts were normalized using DEseq2 and pair-wise comparisons were performed.