Epz 6438

EPZ-6438 (A8221, ApexBio Technology) was dissolved in DMSO and then diluted with 5% sucrose solution as vehicle. The virgin males and females were fed with various doses of EPZ-6438 for 7 days after eclosion to inhibit E(z) enzymatic function in the F0 parents only. The doses of 100, 200 and 400μM were first used to determine an effective dose with western blotting at 100uM (data not shown), and then doses of 50, 100 and 150μM were used for longevity and further western analyses. EPZ-6438 was also mixed into the LP diet or CD at a concentration of 150μm for longevity analyses (see Figures 4–5 for more information).

GBM (U-87) cell line was obtained from NCCS, Pune. Inhibitor PI-103 was procured from Echelon Biosciences and EPZ-6438 from APExBIO. CD24 antibody, Fluo-4 AM, DCFDA/H2DCFDA and Oregon green 488 were procured from Invitrogen. MENA antibody was purchased from Novus, USA. A2BR antibody was procured from Alomone labs, Israel. Phalloidin- Actin was obtained from Cell signaling. α-tubulin antibody was procured from sigma. β-catenin, SMAD1, SNAI3 were procured from cloud clone (USA). All the media were purchased from HiMEDIA, India.

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.