GSM8155374: CD45+ sorted tumor, control treated, day 21 VDJ; Mus musculus; OTHER
For single-cell RNA sequencing of intratumoral immune cells, 4-6-weeks-old C3H/HeJ mice were injected subcutaneously into the right flank with 1x106 SW1 melanoma cells. After 10 days, a pretreatment mouse cohort (n=3) was sacrificed, and the harvested tumors were subjected to scRNAseq as the day 0 of L-fuc treatment timepoint. The remaining mice were administered either control (n=6) or 100nM L-fuc (n=6)-supplemented water ad libitum as previously described(Lau et al., 2015;Lester et al., 2023) for 7 or 21 days (tumor endpoint for harvest). Upon sacrifice, tumors were removed and CD45+ cells were isolated through sorting. Tumors were harvested at 0, 7 or 21 days after the start of L-fuc treatment, and subjected to scRNAseq analyses by 10X Genomics Chromium Single Cell Controller for single-cell RNA-sequencing library preparation (10X Genomics, Pleasanton, CA). Reverse transcription was then performed on encapsulated individual cell droplets to generate cDNA and generate cDNA libraries. These libraries were sequenced using an Illumina NextSeq 500 instrument using v2.5 flow cells; a 10X Genomics CellRanger software was used for alignment and gene counting.