Western Blotting Protocol for Akt Signaling

Primary antibodies used for Western blotting: anti-Akt-1 (1:1000 dilution, cat.# 4691, Cell Signaling); anti-phospho-Akt-1 (S473) (1:1000 dilution, cat.# 9018, Cell Signaling); anti-phospho-Akt-1 (T308) (1:1000 dilution, cat. 13038, Cell Signaling); anti-β-tubulin (1:1000 dilution, cat. 2128, Cell Signaling); anti-ARID1A (1:1000 dilution, cat.# NB100-55334, from Novus Biologicals). PARPi were purchased: ABT-888 from Enzo Life Sciences (cat.# ALX-270-444-M001); Olaparib from Selleck Chemical LLC (cat.# S1060). An inhibitor of PI3K LY294002 was obtained from Selleck Chemical LLC (cat.# S1105) and allosteric Akt inhibitor MK-2206 from Cayman Chemical (cat.# 11593). ARID1A Silencer™ siRNA and Silencer™ Select Negative siRNA Control were purchased from ThermoFisher Scientific (cat.# 4392420 and 4390843). Lipofectamine RNAiMAX (cat.# 13778075, ThermoFisher Scientific) was used for siRNA transfection according to manufacturer recommendations.