The pore-forming apolipoprotein APOL7C drives phagosomal rupture and antigen cross-presentation by dendritic cells

Splenic dendritic cells were isolated and processed using 10X genomics 3' gene expression kit v3.1, to assess APOL7C expression and response to poly(I:C) Overall design: In the first experiment, a 1:1 ratio of panDC and CD45 positive cells, from the spleen were suspended in 50 ul of 1.0% BSA. In the second experiment, lineage (TER-119, CD19, B220, TCRB, NK1.1, Siglec-f, F4/80) -negative, FLT3+ cells from the spleen were suspended in 50 ul of 1.0% BSA. All samples were processed according to 10X Genomics ChromiumTM Single Cell 3' Reagent Guidelines Next Gem Chemistry as per the manufacturer's protocol. In brief, cultured cells were partitioned into Gel Bead-In-EMulsions (GEMs) using 10X Genomics 3' Gene Expression v3.1 technology. Quality control and cDNA quantification was performed using DTape station 1000. Sequencing was performed using Illumina NovaSeq with 100 cycle flow cells. We recovered around 8,500 cells / Sample, with a targeted sequencing depth of ~20,000 reads/cell.