Upon murine necropsy, ascites, splenocytes, and bone marrow were collected and processed into single-cell suspensions through filtration and stored in freezing media (10% DMSO in FBS, Atlas Biologicals) at −80C. All single-cell suspensions were thawed on ice, washed once with 2% BSA in PBS and stained with live/dead UV stain (Invitrogen), and subsequently blocked in FACS buffer (PBS, 2% BSA) containing FcR blocking reagent at 1:50 (Miltenyi Biotec) for 15 minutes. After live/dead staining and blocking, antibody cocktails for myeloid or lymphoid panels (Supplemental Table 2 ) were incubated with cells at a 1:50 dilution for 20 minutes on ice before being washed and suspended in FACS buffer for analysis.
Cell populations were analyzed using an LSRFortessa (BD Biosciences) and were separated and quantified using FlowJo software (RRID:SCR_018933; Tree Star Inc.). Gating methods for myeloid and lymphoid populations were performed following standardized gating strategies previously described (32 ). For complete gating strategies, please seeSupplemental Figure 1 .
Cell populations were analyzed using an LSRFortessa (BD Biosciences) and were separated and quantified using FlowJo software (RRID:SCR_018933; Tree Star Inc.). Gating methods for myeloid and lymphoid populations were performed following standardized gating strategies previously described (32 ). For complete gating strategies, please see