Fig 1: Immunophenotyping human and mouse-derived eGFP-transduced cancer cells by flow cytometry.A. Gating strategy to analyze the reprograming efficiency of mouse-derived eGFP-transduced cancer cells and B. human-derived eGFP-transduced cancer cells at day 9 post induction of PU.1, IRF8, and BATF3 (PIB). Live cells were gated on viability dye negative population (4,6-diamidino-2-phenylindole, DAPI) followed by singlet selection. Expression of CD45+MHC-II+ was quantified by flow cytometry following a gating strategy based on fluorescence minus one (FMO) staining. eGFP-transduced cells were used as controls. C. Mouse and D. human tumor-APCs were evaluated by flow cytometry for the expression of PD-L1, ICOSL, TIM-3, and VISTA at day 9 of reprogramming. The gating strategy was based on FMO for the respective fluorochrome. eGFP-transduced cancer cells and splenic (mouse, live, lineage negative for B220, CD3, NK1.1, CD19, and Gr1, MHC-II+CD11c+CD8a+) or peripheral blood (human, live, lineage negative for B220, CD3, CD56, and CD19, HLADR+CD11c+CD141+) cDC1 cells were used as controls (biological replicates n = 3–11). Mean ± SD is represented.