Vemurafenib Cytotoxicity in Melanoma Cells

SK-mel 28 and SK-mel 24 cells were grown in Gibco Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific, 11965–092), supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin and incubated at 37°C in 5% CO₂. Cell number was measured using a Cellometer Mini Cell Counter (Nexcelom Bioscience); the cell suspension was mixed at a 1:1 ratio with the trypan blue solution before the measurement. For all BRAFi experiments, SK-mel 28 and SK-mel 24 cells were treated with a 2 μM dose of vemurafenib, based on previous literature.^{22 (link)} For HP microcoil experiments, cells were plated 24 hours before treatment and incubated overnight to ensure cell adherence. SK-mel 28 and SK-mel 24 cells were treated with either vemurafenib or 0.1% dimethyl sulfoxide (DMSO, vehicle) for 24 or 48 hours. Immediately before the dissolution of HP [1-¹³C] pyruvate, cells were trypsinized, counted and divided into cell pellets of 2 × 10⁶ per sample. The order of samples loaded using the microcoil multi-shot protocol (Figure 1B) alternated between vemurafenib and vehicle treatment.