GSM7050011: CD45+cells from E0771 tumor of FLT3L-treated mice; Mus musculus; RNA-Seq
The extraction protocol can be found in the author's manuscript: The regular tissue processing procedure was followed, with the addition of actinomycin D (Sigma-Aldrich, A1410-5MG) to each buffer. Tumor collection was performed in 30 µM, enzyme incubation and subsequent filtering in 15 µM, and all other steps in 3 µM. 10^6 cells were stained with anti-CD45 APC-Cy7 (1:50, Biolegend), a mouse cell surface protein antibody panel containing 167 oligo-conjugated antibodies for cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) and TruStain FcX™ PLUS Antibody (1:25; Biolegend) in Phosphate buffered saline (PBS) with 0.04% bovine serum albumin (BSA) for 30 minutes on ice. After incubation, cells were incubated with 7AAD (1:100) 5 minutes prior to sorting. 8x10^4 living CD45+ cells were sorted using BD FACSAriaTM II and collected in 500 μl PBS + 0.04% BSA and 3 μM ActD. The library construction protocol was described in Antunes et al., Nat Neurosci; 2021;24(4):595-610. doi: 10.1038/s41593-020-00789-y. scRNAseq 10x v3.1