Anti phospho akt ser473 and thr308

Cells and reagents. U-2932 (DLBC) and Granta-519 (MCL) cell lines were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ; Braunschweig, Germany). SUDHL-10 (DLBC) cell line was from the American Type Culture Collection (ATCC; Rockville, MD, USA). Cells were maintained in RPMI-1640 medium (Mediatech, Herndon, VA, USA) supplemented with 10% fetal bovine serum (FBS), 2 mM sodium pyruvate and 100 units/ml penicillin/streptomycin at 37˚C in a humidified atmosphere containing 5% CO 2 . The doubling time of each of these three cell lines is ~48 h. PF-04691502 (PI3K(α/β/δ/γ)/ mTOR dual inhibitor), CAL-101 (p110δ inhibitor) and INK-128 (mTORC1/2 inhibitor) were purchased from Selleckchem (Houston, TX, USA). The compounds were dissolved at 50 mM in DMSO as a stock solution, and then further diluted to desired concentrations for in vitro experiments. Rituximab was a kind donation by the affiliated hospital clinic. Antiphospho-Akt (Ser473 and Thr308; 1:500 dilution), anti-Akt (1:500 dilution), anti-phospho-S6 (1:1,000 dilution) and anti-GAPDH (14C10; 1:1,000 dilution) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-PARP (H-250; 1:500 dilution) and anti-cyclin D1 (1:500 dilution) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).