Culturing HEK293 and Uveal Melanoma Cells

HEK293 cells were cultured in Dulbecco's modified Eagle's medium (D6429; Sigma–Aldrich, Inc) containing 10% fetal bovine serum (F2442; Sigma—ldrich, Inc), 1× antibiotic/antimycotic solution (A5955; Sigma–Aldrich, Inc), and 1× Plasmocin prophylactic (ant-mpp; InvivoGen). HEK293 Gαq/11 KO cells were cultured using the same conditions described previously and were a kind gift from Dr Asuka Inoue (41 (link)). UM cells (92.1, OMM1.3) were cultured in RPMI1640 (R8758; Sigma–Aldrich, Inc) containing 10% fetal bovine serum (F2442; Sigma–Aldrich, Inc), 1× antibiotic/antimycotic solution (A5955; Sigma–Aldrich, Inc), and 1× Plasmocin prophylactic (ant-mpp; InvivoGen). All cell lines were routinely tested free of mycoplasma contamination. VS-4718 (S7653), BYL719 (S2814), TGX221 (S1169), CAL101 (S2226), and BKM120 (S2247) were purchased from SelleckChem. FR900359 was prepared in the laboratory of Dr Evi Kostenis. CNO (4936) was purchased from Tocris, Inc EGF (E9644) was purchased from Sigma–Aldrich, Inc. All compounds were used at concentrations indicated in figure legends.