LLC cells exhibit a blunted response to IFNγ in vitro and in vivo compared with CMT167. LLC cells exhibit a blunted response to IFNγ in vitro and in vivo compared with CMT167. CMT167 or LLC cells were orthotopically injected into the left lung lobe of transgenic GFP-expressing C57BL/6J mice and were grown for either 2 (LLC) or 3 wk (CMT167). Tumor-bearing lung lobes were isolated and made into single-cell suspensions containing both GFP-positive (host cells) and GFP-negative (cancer cells). First, RNA was isolated from identical cancer cells grown in passage (in vitro condition). Second, RNA was isolated from recovered GFP-negative cancer cells (isolated via FACS-in vivo condition). Third, RNA was run for RNA-Seq from both conditions. The CMT167 condition had three experimental replicates per in vitro and in vivo conditions with five tumor-bearing lung lobes pooled per in vivo experimental replicate (15 mice used total). The LLC condition had five experimental replicates per in vitro and in vivo conditions with four tumor-bearing lung lobes pooled per in vivo experimental replicate (20 mice used total). (A) The “HM_IFNg Response” pathway containing 80 genes from the Broad Institute Molecular Signatures Database was used to generate a heat map of differentially expressed genes between the LLC and CMT167 in vitro and in vivo experimental conditions. Source data are available for this figure. Bonnie L Bullock et al. LSA 2019;2:e201900328 © 2019 Bullock et al.