Cisplatin

Tumor growth curves of the cSCC-PDX were documented by the dynamic measurement of tumor volume. The tumor volume range of 300–400 mm³ in the tumor-bearing NOD/ShiJic-scidJcl mice was randomly assigned, and a treatment protocol initiated. Each treatment group had a minimum of n = 3 mice per condition. The control (n = 3) mice were intravenously injected with 150 μL of PBS once per week. In the cisplatin group, cisplatin (n = 4, 5 mg/kg, FUJIFILM Wako, Tokyo, Japan) was administered intraperitoneally once per week as described by Lu et al.^{62 (link)}. Eribulin (n = 4) was administered intravenously once per week^{63 (link)}. The tumor volume was measured twice per week by caliper, and tumor weights were measured at 42 days after treatment initiation. Tumor volume and weight were recorded in a blinded manner.

Methyl methanesulfonate (MMS) and thiabendazole (TBZ) were purchased from Sigma-Aldrich (St Louis, MO). Camptothecin (CPT), hydroxyurea (HU), doxorubicin and cisplatin were from Wako Pure Chemical Industries Ltd (Osaka, Japan). Vorinostat/suberoylanilide hydroxamic acid (SAHA) was synthesized in-house (see Supplementary Materials)⁶⁹. doxorubicin was dissolved in water, SAHA (refer supplementary procedure) in DMSO, and cisplatin (Wako, Pure Chemical Industries, Ltd, Osaka, Japan) in 10% NaCl. All drugs were further diluted with their respective solvents prior to use according to manufacturers’ recommendations.

Cisplatin (cis-diamminedichloro-platinum(II): CDDP) and 5-FU were purchased from Wako (Osaka, Japan). Imatinib was purchased from Focus Biomolecules (Plymouth Meeting, PA, USA).

Human fibrosarcoma cell line HT1080 was grown in Dulbecco’s Modified Eagle Medium (DMEM), 10% heat-inactivated fetal bovine serum (FBS), and 1% penicillin–streptomycin solution, at 37 °C under a 5% CO₂ atmosphere. All reagents were obtained from commercial sources: cisplatin (CDDP), N-acetylcysteine (NAC), 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) (WAKO, Osaka, Japan), propyl gallate (Sigma, St. Louis, MO, USA), and Z-VAD-fmk (peptide institute, Osaka, Japan). The antibodies used were against FLAG (Sigma), LKB1, total AMPKα, caspase-3 (Santa Cruz, Dallas, USA), phospho-p38 (threonine 180 and tyrosine 182), total p38, phospho-JNK (threonine 183 and tyrosine 185), total JNK, p53 (Cell signaling technology, Danvers, USA), and β-actin (Wako).