Fig. 4From: Inflammatory cytokines and organ dysfunction associate with the aberrant DNA methylome of monocytes in sepsisIn vitro exposure to LPS induces the acquisition of tolerance and DNA methylation changes similar to those observed in sepsis. a Schematic diagram depicting our in vitro model for sepsis. PBMCs were cultured with or without LPS during 4 days, and then, monocytes were sorted as CD14+CD66b− cells for subsequent analyses. In parallel, monocytes were isolated with magnetic CD14 antibody and cultured in the same conditions. b TNFα and IL-10 production, as determined by ELISA, from PBMCs or monocyte supernatants, following washout and resting following the 4 initial days (with/without LPS) and an 8-h exposure to LPS, as indicated in the scheme in a. c Bisulfite pyrosequencing of selected hypermethylated (IL15) and hypomethylated (IL1R2) genes in sorted CD14+CD66b− monocytes from PBMCs in the in vitro sepsis model. d IL1B, IL6, IL15, and IL1R2 mRNA levels were analyzed by quantitative RT-PCR using RPL38 as control in the same sorted monocytes. e Bisulfite pyrosequencing of IL15 and IL1R2 in CD14+ monocytes in the in vitro sepsis model. f IL1B, IL6, IL15, and IL1R2 mRNA levels analyzed by quantitative RT-PCR in CD14+ monocytesBack to article page