(A-C) A375 WT and CRT cells (CRT34 and CRT35) were treated with PLX4720 (1 μmol/L) and PD0325901 (35 nmol/L) for 48 hours and with raptinal 5 μM for 3 hours. YUMM1.7 WT and CRT cells (CRT47R and CRT49N) were treated with PLX4720 (1 μmol/L) and PD0325901 (35 nmol/L) for 24 hours and with raptinal 2.5 μM for 3 hours. (A) Levels of GSDME, cleaved caspase-3 and PARP in lysates, or HMGB1, IL-1α and IL-1β in supernatants. Vinculin, α-tubulin or ponceau red were utilized for loading controls. Western Blots shown are representative of three independent experiments. (B) Percentage of LDH release representative of two independent experiments. Three replicate wells were performed for each experimental condition. Error bars are SEM. (C) Cell morphology visualized with the IncuCyte S3 system at x10 objective. Plasma membrane bubble-like protrusions, a characteristic feature of pyroptosis, are indicated by red arrow. Scale bar: 100 μm. Cell morphology shown are representative of at least two independent experiments.