Bulk RNA-Seq of Bone Marrow-Derived Macrophages Exposed to Avirulent Coccidioides posadasii (?cts2/?ard1/?cts3)
To verify that Spp1+ macrophage differentiation is contact dependent, macrophages were exposed to Coccidioides in vitro directly, or indirectly via conditioned media (cm), and expression of the gene signature was evaluated by RNA-seq. Overall design: First, we differentiated macrophages with each cytokine and subsequently exposed them to Coccidioides in vitro. GM-CSF differentiated bone marrow-derived macrophages (GM-BMDM) and M-CSF differentiated macrophages (M-BMDM) were incubated overnight with Coccidioides arthroconidia. Next, to determine if Spp1 induction was contact-dependent, fresh GM-BMDM were exposed to CM collected from control GM-BMDM or GM-BMDM incubated with Coccidioides (GM-BMDM + cocci). RNA-seq was conducted on RNA isolated from the following groups: GM-BMDM, GM-BMDM + CM, GM-BMDM + cocci, and GM-BMDM + cocci CM.