Western Blot Analysis of Cell Signaling

Following total protein extraction, western blotting was performed as previously described [30 (link)]. Primary antibodies used in this study were as follows: rabbit anti-Akt (Cat. # 9272), pAkt (S473, Cat. # 9271), LKB1 (Cat. # D60C5) and PARP (Cat. # 9542) antibodies were from Cell Signaling Technology (Danvers, MA), mouse anti-FAK (clone 77/FAK) was from BD Transduction Laboratories (Mississauga, ON), and mouse anti-β-actin antibody (clone AC-74) was from Sigma (St. Louis, MO). Following incubation with primary antibody overnight, membranes were washed 3 x 5 minutes and incubated in horse radish peroxidase (HRP) conjugated goat anti-mouse or goat anti-rabbit secondary antibody (Calbiochem, San Diego, CA) for 1 hour. Membranes were then washed 6 x 5 minutes and incubated in Immobilon Western Chemiluminescent HRP Substrate (EMD Millipore, Billerica, MA) prior to image development using the GeneGnome Bio Imaging System and GeneSnap software (Syngene, Frederick, MD).