Fig 1: Generation of MDDCs for HLA-II immunopeptidomic profiling(A) Schematic depicting the workflow used to generate the recombinant S protein pulsed MDDC immunopeptidome. MDDCs were pulsed with S or control protein for 18 h, and sequential HLA IPs were carried out as indicated. Peptides were eluted from HLA molecules, purified by preparative HPLC, and subjected to a LC-MS/MS identification workflow.(B) MDDCs were harvested following overnight protein pulse and stained with antibodies to identify DCs and assess their expression of HLA-I, HLA-DR, DC-SIGN, DEC-205, CD83, CD40, CD80, and CD86 and then analyzed by flow cytometry. Histogram plots are gated on live CD11c+CD4+ DCs and show expression of each indicated marker on cells treated with S or control protein. Unstained cells are shown in filled gray histograms. Inset numbers indicate mean fluorescence intensity for each sample.See also Figure S2.