Ly 2584702

The primary antibodies and their dilution ratio in western blot analysis were as follows: p-mTOR-S2448 (1:1,000, Cell Signaling Technology); mTOR (1:1,000, Cell Signaling Technology); p-p70S6K-T389 (1:1,000, Cell Signaling Technology); p-p70S6K-T421/S424 (1:1,000, Cell Signaling Technology); p70S6K (1:1,000, Cell Signaling Technology); p-S6 Ribosomal Protein -S235/S236 (1:1,000, Cell Signaling Technology); RPS6 (1:1,000,Abcam); β-Actin (1:1,000, Tianjin Sungene Biotech Co., Ltd., China) and GAPDH (1:1,000, Good Here, China). DHA was purchased from Tokyo Chemical Industry (Cat #D3793, Japan). MK-2206 (AKT inhibitor, Cat #A10003) was purchased from Selleckchem. LY2584702 (p70S6K inhibitor, Cat #1082949–68–5) was purchased from Med Chem Express.

SCC25 (1 × 10⁶ cells/mouse) or Cal27 (1 × 10⁶ cells/mouse) were inoculated subcutaneously into NSG mice (n = 8 mice per group), and once the volume of the tumors reached 50 mm³, treatments were initiated 6 times a week for up to 4 weeks. Tumor volume was measured twice a week. erlotinib (15 mg/kg), LY (12.5 mg/kg) and TOFA (1 mg/kg) were suspended in aqueous mixture of 0.5% Hydroxypropyl methylcellulose + 0.2% Tween 80 and administered by oral gavage. LY-2584702 was purchused from the MCE (MedChemExpress).
The rest of the drugs were purchased from Cayman chemicals. The Hebrew University is an AAALAC International accredited institute. All experiments were conducted with approval from the Hebrew University Animal Care and Use Committee.