scRNA-seq of CD45+ cells isolated from lungs of mice carrying KPneo tumors that received FLT3L and αCD40 (DC-therapy) or rat IgG2a isotype control
scRNA-seq of CD45+ cells isolated from lungs of mice carrying KPneo tumors that received FLT3L and αCD40 (DC-therapy) or rat IgG2a isotype control
Lungs were harvested at day 9 and digested to obtain single cell suspensions to be processed for scRNA analysis. CD45+ cells were isolated using ARIA Cell Sorter (BD). Single cell suspensions were processed for single cell analysis using Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Dual Index) (10X Genomics) following manufacturer's instructions. 8-weeks females C57/BL6 mice (4 mice per group) were injected with 5x10^4 KPneo cells intravenously. Lungs were harvested at day 9 post-inoculation. Mice were administered 30 ug of recombinant Flt3L on day -2, 2, 4 , 6. On day 5, mice received a single dose of aCD40 (200ug intraperitoneally). Control mice received intraperitoneal injections of PBS on day -2,2,4,6 and isotype rat aIgG on day 6. Nucleic acids (polyA RNA) were extracted during single cell encapsulation and immediately barcoded, according to Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Dual Index) (10X Genomics) protocol. Libraries were constructed using Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Dual Index) (10X Genomics) protocol and following manufacturer's instructions.