Xmarks spectrometer

786-0 and 786-0R cells were seeded in 24 wells plates (Santa Cruz biotechnology, Dallas, Texas) and allowed to attach for 24 hours. Afterwards, cells were treated with increases concentrations of sunitinib (LC laboratories, Woburn, MA), the EZH2 inhibitor GSK126 (Cayman chemical company, Ann Arbor, Michigan) or combination of both sunitinib and GSK126 using combination index values (Calcusyn software). Cell were fixed and stained with crystal violet (Sigma Aldrich, St. Louis, MO) after 48h of treatment. Afterwards, stained cells were washed with ddH₂O to remove excess dye, air-dried and dissolved in methanol. Cells viability was quantitated by absorbance using a spectrometer at 570nm (xMarks Spectrometer, Bio-Rad). 786-0shRNA (scramble) and 786-0shEZH2 were seeded in 24 well plates and 24 hours later, cells were treated with 2μM, 4μM, 6μM and 8μM of sunitinib. After 48 hours of treatment, cells were fixed, stained and read using spectrometer.

786-0 and 786-0R cells were plated in six-well plated for 24 hours and treated with either DMSO (vehicle control) varying concentrations of sunitinib or a single concentration of sunitinib in combination with EPZ011989 (EZH2 inhibitor) and use MTT assay (Cayman) or crystal violet assay (Sigma) to evaluate the cells growth after 24, 48, 72 and 96 hours of treatment and absorbance read using s spectrometer (xMarks Spectrometer, Bio-Rad).