For xenograft experiments, female NOD-SCID mice (6 to 8 weeks) were subcutaneously injected with 3 × 106 cells per mice in 1:1 Matrigel (Corning). Tumor volumes were monitored by digital calipers twice a week and calculated using the formula: volume = (width)2 × length/2. Trametinib was administered orally (2 mg/kg) every other day when tumors reached an average volume of ~50 to 60 mm3. At the experiment end point, mice were euthanized, and tumors were harvested for ex vivo analysis and subsequent histology. All operations were approved by the Animal Ethics Committee of National Cancer Centre Singapore (NCCS).

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