Capivasertib azd5363

Doxorubicin hydrochloride (#D1515), vincristine, and insulin growth factor 2 (IGF2, #I2526) were purchased from Merck. Irinotecan (#S2217), MK-2206, capivasertib (AZD5363), alpelisib (HY-15244), nilotinib (S1033), pazopanib (S3012), and ponatinib (S1490) were purchased from Selleckchem. Everolimus (RAD001, SRP020750e) was purchased from Sequoia Research Products, etoposide was purchased from Sandoz and AVE 1642 from Immunogen. D-188514, an ifosfamide analog that does not require metabolic activation, was purchased from Niomech. The PI3K/mTOR dual inhibitor NVP-BEZ235 was kindly provided by Novartis. Trabectedin (ET-743) was kindly provided by PharmaMar. Working dilutions of all drugs were prepared immediately before use.

E2A-PBX1⁺ B-ALL cell lines RCH-ACV and 697, as well as E2A-PBX1^- B-ALL cell lines REH and SEM were obtained from DSMZ (Braunschweig, Germany) in 2013, and were authenticated in 2020 in DSMZ. They were monitored to exclude mycoplasma contamination regularly. Culturing of human cells was performed as previously described [7 (link)]. RPMI-1640, IMDM, DMEM, and Opti-MEM mediums were obtained from Thermo Fisher (Waltham, MA, USA). Torin-1, everolimus (RAD001), ipatasertib (GDC-0068), alpelisib (BYL719), capivasertib (AZD5363) and idelalisib (CAL-101) were from Selleck Chemicals (Houston, TX, USA). FBS was from Sigma-Aldrich (St. Louis, WIS, USA).

Short hairpin RNA (shRNA) oligonucleotides targeting Slc6a8, p65/NF-κB, HIF1A and HIF2A, and control shRNA-NC were all purchased from GenePharma (Shanghai, China). The breast cancer cells (MDA-MB-231 and BT-549) that stably expressed above shRNAs were established by lentivirus transduction according to the manufacturer’s instructions. The sequences of shRNA used were listed in Supplementary Table 1. The promoter containing TP53/FOS/ETV4/p65/NF-κB-wild type binding sites (WT) or mutated binding sites (MUT) was cloned into pGL3 luciferase reporter vector to obtain the pGL3/Slc6a8 WT reporter and pGL3/Slc6a8 MUT reporter (GenePharma, China). The AKT inhibitor Capivasertib (AZD5363) was purchased from Selleckchem (USA).