S2827

Torin 1 (S2827, Selleckchem), PD98059 (A1663, ApexBio) and danusertib (A4116, ApexBio) were injected via intraperitoneal (i.p.) injection, daily, at the dosage of 6.8, 10 and 15 mg kg⁻¹, respectively. EPZ6438 (A8221, ApexBio) was injected via i.p. injection once every 2 days at 50 mg kg⁻¹. For antibody therapy, 4 mg kg⁻¹ DECMA-1 (U3254; Sigma) or rat immunoglobulin G (I4131; Sigma) was injected via i.p.injection twice a week. In the experiments using MCF-7 cells, oestradiol pellets were prepared and transplanted into nude mice before cancer cell injection, according to a previously published protocol50 (link), unless otherwise noted.

Endemic Burkitt lymphoma cell lines HH514-16 (a gift from Dr. George Miller, Yale University), Jijoye and Raji (gifts from Dr. Janet Hearing, Stony Brook University) were maintained in RPMI 1640 supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco). CLIX-FZ (Clone-HH514-16 transfected with pLIX_402-FZ) cells were generated from puromycin (P8833, Sigma-Aldrich) selected HH514-16 cells transfected with pLIX_402-FZ. Lymphoblastoid cell line (LCL) used was generated and maintained as described before [40 (link)]. Sodium butyrate (NaB; 3mM; 303410, Sigma-Aldrich), doxycycline (5μg/ml; D9891, Sigma-Aldrich), KU-55933 (1μM; SML1109, Sigma-Aldrich), Torin1 (0.5μM; S2827, Selleckchem) and chloroquine (200μM; C6628, Sigma-Aldrich) were used to treat Burkitt lymphoma cells except where specific concentrations are indicated. KU-55933 (1μM) and chloroquine (100μM) were used to treat lymphoblastoid cells except where specific concentrations are indicated.

The tails were amputated at the 6th caudal vertebrae to set up the regeneration models. Akt inhibitor MK-2206 (5 mg/kg body weight (BW) S1078, Selleck, Houston, TX) was administered after autotomy by i.p. injection every other day. BrdU solvent (20 μL of 20 mM solution, B5002, Sigma, Darmstadt, Germany) was administered on the last 2 days before sample collection. The samples were collected on the 7th day or other time point indicated. For the treatment of mTOR inhibitor (RAD001, Torin1) and AMPK activator (A769662, Selleck, Houston, TX), the gecko models were administered RAD001 (10 mg/kg BW, S1120, Selleck, Houston, TX), Torin1 (20 mg/kg BW, S2827, Selleck, Houston, TX) and A769662 (30 mg/kg BW, S2697, Selleck, Houston, TX) individually or together by i.p. injection post autotomy every other day. For the treatment of Wnt agonist, CHIR98014 was administered (5 mg/kg BW, S2745, Selleck, Houston, TX) by i.p. injection post autotomy every other day. The BrdU was administered as mentioned above. For treatment with the Wnt inhibitor, IWP-4 (2 mg/kg BW, SML1114, Sigma, Darmstadt, Germany) or XAV939 solution (40 mg/kg BW, S1180, Selleck, Houston, TX) was administered after amputation by i.p. injection every other day.