Multicellular melanoma spheroids were cultured as described.21 (link) Briefly, the 96-well plate was precoated with 50 µL 1.5% agarose before seeding CFSE (Thermo Fisher) labeled A2058 melanoma cells at 2×104 cells/well and allowed to form spheroids over 48 hours. After coculturing with γδ T cells, an annexin V/propidium iodide (PI) or PI (Biolegend) staining was performed for further analysis by flow cytometry or confocal microscope, respectively. The supernatant was harvested for cytotoxicity by LDH releasing assay (Promega) as described earlier.
Single-cell RNA sequencing (scRNA-seq) dataset scRNA-seq data of sorted γδ (Vδ2) T cells from three healthy human donor PBMCs were published by Pizzolato et al22 (link) and obtained from NCBI GEO data set repository (GSE128223). Three preprocessed mRNA datasets of GSM3667468 (763 cells), GSM3667470 (1277 cells), and GSM3667472 (1720 cells) were downloaded and analyzed with pipelines provided by Seurat Package.23 (link)
Single-cell RNA sequencing (scRNA-seq) dataset scRNA-seq data of sorted γδ (Vδ2) T cells from three healthy human donor PBMCs were published by Pizzolato et al22 (link) and obtained from NCBI GEO data set repository (GSE128223). Three preprocessed mRNA datasets of GSM3667468 (763 cells), GSM3667470 (1277 cells), and GSM3667472 (1720 cells) were downloaded and analyzed with pipelines provided by Seurat Package.23 (link)
