Socs1 KD tumors show alterations in multiple populations.

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Socs1 KD tumors show alterations in multiple populations. Socs1 KD tumors show alterations in multiple populations. LLC-NT or LLC-sh21 cells were orthotopically injected into the left lung lobes of mice and established primary tumors. After 2 wk of tumor growth with no treatment, mice were euthanized and their tumor-bearing lung lobes were isolated. Single-cell suspensions were made from tumor-bearing lung lobes or naive lungs. Naive biological replicates each contained lungs isolated from one mouse (three mice/three replicates total). LLC-NT and LLC-sh21 biological replicates each contained tumor-bearing lungs from three pooled mice (nine mice/three replicates total; three mice/replicate). Single-cell suspensions were then stained with a 39-antibody panel and analyzed on the Helios mass cytometer. Data show all viable single cells, subjected to the PhenoGraph algorithm. (A) Primary tumor volume was assessed by digital calipers. (B) PhenoGraph-based visualization on a tSNE plot, colored according to the expression of the cellular markers (parameters): CD3, CD4, CD8, CD11b, CD11c, CD19, CD44, CD45, CD64, CD90.2, Gr-1, MHC-II, and Siglec-F. (C) PhenoGraph-defined cellular distribution and clustering, as defined by tSNE1 and tSNE2, colored by cluster ID for all experimental groups and replicates. (D) Pie charts show the abundance of all clusters containing cancer cells (clusters #5 and #35) identified as CD45−and missing in naïve samples. (E) Cancer cell clusters in LLC-NT samples or LLC-sh21 samples colored according to Ki67, MHCI, MHCII, and PD-L1. Bonnie L Bullock et al. LSA 2019;2:e201900328 © 2019 Bullock et al.