MTT assay

Pazopanib was purchased from Adooq Bioscience (GW-786034). Pazopanib was diluted in dimethyl sulfoxide to 10 mM and the required concentrations were added to the respective media.

Using a 96-well plate, the following cell numbers of each sarcoma cell line were seeded in individuals wells: 2,500 of SW872, 500 of HT1080, 3,000 of SK-LMS-1, 2,000 of A204, 2,500 of RD, 2,000 of ISO-HAS-B, 1,000 of S462, 2,000 of FMS-1, 1,000 of SFT8611, 2,500 of SFT9817, 2,500 of HS-sch2, 3,000 of YST-1, 2,000 of HS-SY-II, 3,000 of Yamato-SS, 2,000 of SaOs2, and 1,000 of MG-63. To calculate the pazopanib inhibitory potency (IC₅₀) for the 16 sarcoma cell lines, at least seven different doses of pazopanib were used. Cells were treated with pazopanib and were seeded 24 h after onto the plates. The cell viability was measured after 72 h using the Cell Counting Kit-8 (Dojindo Laboratories), according to the manufacturer's protocol. Cell viability (%) was calculated by dividing the median value for each pazopanib dose (conducted at least in triplicates) by the value of untreated cells. IC₅₀ values were calculated using the dosages from the two-cell viability values surrounding 50%. From the results obtained from three or more independent experiments, we calculated the IC₅₀ data.