title: GSM8616656: DC-1_r1_ADT; Mus musculus; OTHER
library_construction_protocol: 21 or 42 days following vaccination, popliteal lymph nodes were harvested from 13 mice, with 4 lymph nodes mechanically separated with 22-gauge needles in 2 mL of EHAA media. 8 LNs were digested in collagenase D and DNase 1 for isolation of dendritic cells. The other 18 LNs were digested with Liberase DL with DNase 1 to isolate the stromal cell population. The 8 LNs digested for dendritic cell isolation were then stained with CD11c, CD11b and B220, and a live/dead dye. Live cells were then sorted into four tubes on a FACS Aria Cell Sorter: sorted CD11c-APC Cy7+ cells, sorted CD11b PE-Cy7+ cells, sorted B220 PE+ cells and Fixable Viability Stain 510 ungated live cells, which were recombined at a 4:4:1:1 ratio, respectively. For the remaining lymph nodes in the stromal cell digest, cells were stained with CD45-PE and Ter119-PE followed by magnetic bead isolation using the Miltenyi bead isolation kit. CD45-negative cells that passed through the column were then washed and counted using a hemocytometer. Cells were assayed using the 10x Genomics v3 single-cell 3' expression kit or v3 single-cell 3' HT kit according to manufacturer's instructions and CITE-seq protocol.