The SVT2 were seeded on six-well plates at a density of 2.5 × 105 cells/well, whereas A431 was seeded at a density of 3 × 105 cells/well. After 24 h, cells were treated with 9.8 μM of 1-Me,Me or 2.3 μM 1-Oct,Me . After 48 h of incubation, Western blot analyses were performed by using pro-caspase 9 (Cell Signaling Technology, Danvers, MA, USA) and pro-caspase 3 (Abcam, Cambridge, MA, USA) antibodies, as reported by Del Giudice et al. [48 (link)] Protein intensity levels were normalized using β-actin (Sigma-Aldrich, St. Louis, MO, USA). The chemiluminescence detection system was purchased from Bio-Rad (Hercules, CA, USA).
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