Mst2 smartpool sirna

Cell culture and transfection: A375, SK-Mel28, WM-793 and SK-Mel2 were obtained from ATCC and were validated by sequencing before initiation of the study. All cells were grown in RPMI media (Gibco, USA) supplemented with 10% foetal bovine serum (Gibco) and 2 mM L-glutamine (Gibco). Media for the resistant cells contains 2 μM PLX4032 (Selleck chemicals, USA). HeLa cells were also validated by sequencing and grown in DMEM (Gibco) 10% foetal bovine serum and 2 mM L-glutamine. The cells were maintained at 37°C and 5% CO 2 . Cells were transfected with Lipofectamine 2000 (Invitrogen, USA) following manufacturers protocol and the amount of DNA and siRNA is indicated in each experiment. FLAG-BRAF, -BRAF R509H , -BRAF V600E/R509H , and BRAF V600E , MYC-LATS1 and FLAG-MST2 have been described (Romano et al., 2014a , Quinn et al., 2021 , Jambrina et al., 2016) HA-Ubiquitin was a gift from Edward Yeh (Addgene plasmid # 18712) (Kamitani et al., 1997) . MST2 SMARTpool siRNA is from Dharmacon (M-012200, CO, USA) and was validated before (Matallanas et al., 2007) . Bortezomib (S1013), trametinib (S2673) are from Selleck Chemicals and Proteasome inhibitor I from Calbiochem (539160, CA, USA).