P akt thr308

Antibodies used for immunoblots include Rheb (Cell signalling, Cat# 1387, 1:1000), Cre (Cell signalling, Cat# 12830, IHC/IHF - 1:50), Cleaved Caspase 3 (Cell signaling, Cat# 9661, IHC - 1:500), pAkt S473 (Cell signaling, Cat# 4060, 1:1000), pAkt Thr308 (Cell signaling, Cat# 13038, 1:1000), pS6 S240/244 (Cell signaling, Cat# 5364, WB - 1:1000, IHC/IHF – 1:500), Total S6 (Cell signaling, Cat# 2317), p4EBP1 S65 (Cell signaling, Cat# 13443, WB-1:1000, IHC/IHF-1:500), p4EBP1 Thr37/46 (Cell signaling, Cat# 2855, WB-1:1000, IHC/IHF-1:500), 4EBP1 (Cell signaling, Cat# 9644, WB-1:1000), eIF4E (Cell signaling, Cat# 9742, 1:1000), Cyclin D1 (Cell signaling, Cat# 2978, 1:1000), Cyclin D1 (Cell signaling, Cat# 2922, IP-1:50), Cyclin D3 (Cell signaling, Cat# 2936, 1:1000), Actin (Sigma, A5316, 1:5000), PyV mT (clone 762, a gift from Dr. Steven Dilworth, IHC/IHF-1:100), Akt1/2 (Santa Cruz, SC-1619, 1:200), E-Cadherin (BD Bioscience, Cat# 610182, 1:1000), Neu (Dako, A0485, 1:500), Ki67 (Abcam, ab15580, 1:500), Cytokeratin 8/18 (Fitzgerald, 20R-CP004, 1:500), Cytokeratin 14 (BioLegend, PRB-155P, 1:500), and Anti-puromycin (Millipore, Clone 12010 – MABE343, 1:1000). Goat anti-rabbit IRDye 800, Goat anti-mouse IRDye 680, Donkey anti-goat IRDye 800 (Li-COR, 925–32211, 925–68070, 925–68074, 1:10 000)

For immunoblot, primary antibodies used were: p-ErbB2 (cell signaling# 2249, 1:1000), ErbB2 (Santa Cruz #284, 1:1000), p110α (cell signaling #4249, 1:1000), p110β (santacruz #602, 1:1000), p110γ (cell signaling #5405, 1:1000), p85 (cell signaling #4257, 1:1000), Pten (cell signaling #9559, 1:1000), p-Akt thr308 (cell signaling #4056, 1:500), p-Akt ser473 (cell signaling #9271, 1:1000), Akt1-2 (santa-cruz #1619, 1:1000), β-actin (Sigma A5441, 1:10000). Secondary antibodies used were conjugated to horserashish peroxidase (HRP) (jackson laboratory). For immunoprecipitation, a p110β antibody (santacruz #602) was used. Primary antibodies for immunoblot were ErbB3 (santacruz #285 1:1000) and p110β (santacruz #602 1:1000)