Bioenergetics analysis in single-cell melanoma suspensions, melanoma cell lines, and HEMs. A. Single-cell suspensions from a ‘fresh’ tumor respond to pharmacologic inhibitors of metabolism, as determined by a Seahorse XF24 Extracellular Flux Analyzer. Depicted are OCR (panel i, left) and ECAR (panel ii, right) rates of single-cell suspensions, prepared from one of four metastatic melanomas. Similar results were obtained with the other 3 tumors. After baseline OCR and ECAR determination, the melanoma cells were treated with oligomycin (O), FCCP (F), rotenone (R), or 2-DG. B. Ratio of basal OCR over basal ECAR in HEMs, melanoma cell lines, and single-cell melanoma suspensions. Data are depicted as the ratio of basal OCR over basal ECAR ratio of short-term cultures of HEMs, six different metastatic human melanoma cell lines (WM983-A, WM983-B, WM1158, WM852, Lu1205, and C32), and single-cell suspensions obtained from four subcutaneous metastatic melanomas.