Cell lines were cultured at 37°C in a humidified atmosphere of 95% air and 5% CO2. All media were supplemented with penicillin (100 U/ml) and streptomycin (100 mg/ml). Fetal calf serum (FCS) (5%) was added to MCDB 153 medium (Sigma) designed for low-serum growth, whereas all other media contained 10% FCS. The UM cell lines 92.1, Mel202 (both carrying mutated GαqQ209L), Mel270 (GαqQ209P-mutated), Mel258, and Mel290 (both Gαq/11wt) were provided by M. Jager (University of Leiden, Netherlands). The metastatic UM cell line OMM1.3 (GαqQ209P) was a gift of B. R. Ksander (Harvard Medical School, USA). All UM cell lines were cultured in RPMI 1640 medium (Life Technologies). The mouse CM cell lines B16 [Gαq/11wt from the American Type Culture Collection (ATCC)] and HCmel12 [with mutated Gα11Q209L generated from a 7,12-dimethylbenz(a)anthracene–induced melanoma from HGF-CDK4(R24C) mice (48)] as well as the human CM cell lines Mamel119 (Gαq/11wt, BRAFwt) and Skmel28 (Gαq/11wt, B-RafV600E) were provided by D. Schadendorf (University of Essen, Germany). The human skin melanoma cell line A375 (carrying B-RafV600E) was obtained from Sigma. CM cell lines were cultured in RPMI 1640 medium complemented with 2 mM L-glutamine, 10 mM nonessential amino acids, 1 mM Hepes (all from Life Technologies), and 20 mM 2-mercaptoethanol (Sigma). HEK293 cells (ATCC) genome-deleted by CRISPR-Cas9 of functional alleles of GNAQ and GNA11 (HEK293 Gαq/11 KO cells) were established as previously described (14). Native and genome-edited HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Life Technologies) with supplements as described earlier.

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