Fig. 4

In 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+ monocytes