Inflammation-produced TNFα protects hematopoietic stem cells from necroptosis and promotes hematopoietic regeneration

To gain molecular insights into the differential effect of TNFα between hematopoietic stem cells (HSC) and downstream myeloid progenitors, we performed RNA-sequencing analyses of HSCs and granulocyte-macrophage progenitors (GMP) treated with TNFα both ex vivo and in vivo. RNA sequencing of 3 biological replicates of HSCs (Lin-/cKit+/Sca1+/Flk2-/CD48-/CD150+) and GMPs (Lin-/cKit+/Sca1-/FcγR+/CD34+) treated ± TNFα both ex vivo for 3 and 12 hours and in vivo for 3 hour (1x3hr). For ex vivo treatment of TNFα, HSCs and GMPs were isolated by FACS from C57BL/6 mouse bone marrow (BM) and cultured in liquid media ± TNFα for either 3 or 12 hours. For in vivo treatment, HSCs and GMPs were isolated from C57BL/6 mice 3 hours after a single injection of PBS or TNFα.