IFN-? induced the formation of foamy macrophages by CD40 signaling control during Mycobacterium abscessus pulmonary infection

Transcriptome sequencing was performed to identify genes that might be responsible for the formation of a large number of lipid droplets with a combination of IFN-? pretreatment and M. abscessus infection Overall design: For transcriptome sequencing analysis of BMDM (2.5×106/well), cells were plated on 6 well glass bottom plate. Media was changed to either BMDM media, or BMDM media with 6.25 ng/mL recombinant murine IFN-? (Novoprotein, Suzhou, China). After overnight IFN-? pretreatment, BMDM were infected with M. abscessus at an MOI of 10. Following the 2-hour phagocytosis period, media was changed and IFN-? was added again at 6.25ng/mL to all IFN-? pre-treated wells. 48 hours post-infection, discard the supernatant and wash twice with sterile PBS. Total RNA was extracted from BMDM cells using TRIzol (Invitrogen, Carlsbad, California, USA) according to manual instruction.