Defining the mechanisms of action to the anti-PD-1+LAG-3 and anti-PD-1+CTLA-4 combinations in melanoma flank models

We used single cell (sc) RNA-seq to analyze responding SM1 melanoma flank tumors and SM1 brain metastases to explore the mechanism of action of the anti-PD-1+LAG-3 and the anti-PD-1+CTLA-4 combination. Tumors were harvested at the endpoint under sterile conditions and weighed. Single-cell suspensions were prepared by enzymatic digestion, using a MACS tumor dissociation kit. The cell count and viability were analyzed by staining the cells with AO/PI stain on the Nexcelom Cellometer K2. Analysis was done using sc-RNAseq.