scRNA-seq of CD45+ cells from mouse B16F10 melanoma treated with anti-PD-1 antibody against untreated control
The B16F10 mouse melanoma cells were inoculated onto the right flank of C57BL/6J mice and every 4 days, anti-PD-1 antibody was injected through the tail vein. In total, 3 doses of immunotherapy treatments were given to the experimental animals. Two days after the last treatment, all tumors including treated and untreated control were collected, cut and digested. The resulted tumor sample single cell suspensions were next red blood cell lysed, stained and then proceeded for FACS sorting to get the live CD45+ intratumoral immune cells. The CD45+ cells from the tumor samples were subsequently count to adjust cell concentration and then used as input for scRNA-seq library construction using the 10x Genomics GEM-X Universal 3' Gene Expression v4 kit following the manufacturer's protocol.
The B16F10 mouse melanoma cells were inoculated onto the right flank of C57BL/6J mice and every 4 days, anti-PD-1 antibody was injected through the tail vein. In total, 3 doses of immunotherapy treatments were given to the experimental animals. Two days after the last treatment, all tumors including treated and untreated control were collected, cut and digested. The resulted tumor sample single cell suspensions were next red blood cell lysed, stained and then proceeded for FACS sorting to get the live CD45+ intratumoral immune cells. The CD45+ cells from the tumor samples were subsequently count to adjust cell concentration and then used as input for scRNA-seq library construction using the 10x Genomics GEM-X Universal 3' Gene Expression v4 kit following the manufacturer's protocol.
European Bioinformatics Institute