Tenovin 1

G361, Hs294T and A375 human melanoma cells and human embryonic kidney HEK293T cells were obtained from American Type Culture Collection (ATCC, USA). G361 cells were maintained in McCoy's 5a medium (Corning Cellgro, USA), and Hs294T, A375, HEK293T in Dulbecco's Modified Eagle's Medium (Corning Cellgro, USA) with 10% FBS (Sigma, USA) at standard cell culture conditions (37°C, 5% CO2 in humidified incubator). Cells were grown to approximately 50% confluence, and treated for 48 h with 10 and/or 25 μM tenovin-1 (Cayman Chemical, USA) dissolved in DMSO. DMSO alone treated cells served as control.

All antibodies were obtained from sources as described in the figure legends. CQ diphosphate salt was obtained from Sigma-Aldrich (St. Louis, MO, USA C6628). CQ diphosphate salt was made up as 100 mM stock in 1× PBS (HyClone, Thermo Scientific, Waltham, MA, USA #SH30256) and freshly made each experiment. Tenovin-1 was purchased from Cayman (Ann Arbor, MI, USA #13085). Vemurafenib was purchased from Selleck Chemicals (Houston, TX, USA #S1267). Vemurafenib stocks were prepared as 40 mM in DMSO (Sigma-Aldrich #D8418) and stored at–20°C. The synthesis of tenovins 6, 30a, 30b, 30d, 30j, 30k, 30n, 33 and 39 was described previously [34 (link)]. The synthesis of tenovins D1 and D3 was described previously [35 (link)]. The synthesis of tenovins 39-OH, 50, 50-OH and 51 is described in supplemental methods. All tenovin compounds were prepared as 40 mM or 60 mM stocks in DMSO (Sigma-Aldrich #D8418) and stored at–20°C.