Next-Generation Standard RNA-Sequencing Analysis of human monocytes stimulated by HMGB1 with or without DMOG

We report the application of standard RNA-sequencing technology for high-throughput profiling of differential gene expression in mammalian cells. This study aims to compare differential gene expression in human monocytes in the absence or presence of HMGB1 and DMOG exposure would mimic hypoxia. Human monocytes (five million cells) from a healthy donor were divided into four groups in triplicate (control, DMOG, HMGB1 or HMGB1_DMOG). RNA isolation, RNA integrity check, library preparation and ~350M raw paired-end reads per lane (Illumina HISeq. 2x150 bp sequencing) were conducted at GENEWIZ, LLC. We find that increased TNF signaling induced by HMGB1 and enhanced when DMOG was present, and reduced IFN signaling in DMOG in unstimulated and HMGB1-stimulated conditions. We conclude that RNA-seq based UP and DOWN regulated genes/pathways characterization would confirm our results and expedite genetic network analyses. Overall design: Examination of different gene expression/signaling pathways in human monocytes from four groups in triplicate (control, DMOG, HMGB1 or HMGB1_DMOG).