NVP-BAG956 (BAG956) is a potent PI3K/PDK inhibitor that inhibits PI3Kδ, PI3Kα, PI3Kγ, and PI3Kβ, shows strong cytotoxicity against T-ALL cell lines, and can be used to study melanoma.

PDK1:240 nM, pAkt:67 nM, A2058 cells:290 nM, VEGFR1:2.56 μM

NVP-BAG956能够抑制PDK1（IC50：240 nM）和VEGFR1（IC50：2.56 μM）。它还可以阻断A2058细胞中PKB/Akt的磷酸化，IC50为67 nM。PKB/Akt磷酸化的抑制与A2058细胞增殖的减少密切相关，NVP-BAG956的IC50为290 nM。在NVP-BAG956的作用下，A2058细胞仅能从G2-M期退出，之后停留在G1期。此外，NVP-BAG956明显诱导A2058细胞中p27Kip1的表达，但对C32细胞没有此作用。[1]

One day after plating (7×10^3 cells/cm2), melanoma cells (A2058, B16F1, B16F10, C32, HBL, Malme, Malme3M, NA8, SKMel2, SKMel23, A375, Hs294T, WM35, and 1205lu cells) are exposed to LY294002 (25 μM), Wortmannin (500 nM), NVP-BAG956 (1 μM), NVP-BBD130 (1 μM), NVP-BEZ235 (1 μM), and ZSTK474 (1 μM), and Rapamycin (100 nM). Compound concentrations are set 2 log units above the IC50 in vitro to ensure full PI3K inhibition, except for the μM inhibitor LY294002. Cells are trypsinized and counted, and the volume is quantified using a Casy Counter and Analyser. To determine the nuclear volume, cells are resuspended in CASYton containing 0.5% Triton X-100, followed by repetitive pipetting (8×), before volume measurements.