Nurd complex antibody sampler kit

Cells and tissues were lysed by homogenization in RIPA lysis buffer (ThermoFisher Scientific, #89900). Protein extracts were separated by SDS-PAGE and then transferred onto PVDF membrane (Millipore, #IPVH00010). Following overnight incubation with the indicated primary antibodies at 4°C, membranes were incubated with IR dye-coupled secondary antibodies (LI-COR) and then visualized by using the LI-COR Odyssey infrared imaging system (LI-COR). The primary antibodies and diluted ratio include: anti-phospho-AKT 1:1000 (Ser473; Cell Signaling Technology, #9271); anti-AKT 1:1000 (Cell Signaling Technology, #2920); anti-β-ACTIN 1:5000 (Sigma, #A1978); anti-IκBα 1:1000 (Cell signaling Technology, #4814); anti-α-Tubulin 1:1000 (Cell Signaling Technology, #3873); anti-Acetylated Lysine 1:1000 (Cell Signaling Technology, #9441); anti-NFκB p65 1:1000 (Cell Signaling Technology, #8242); anti-p300 1:1000 (Cell Signaling Technology, #86377); anti-FLAG (Sigma, #F1804); anti-Acetyl NFκB p65 (Lys310) 1:1000 (Cell Signaling Technology, #12629). NuRD complex components (CHD4, MTA1, HDAC1, RbAP46) antibodies were all from NuRD Complex Antibody Sampler Kit (Cell Signaling Technology, #8349) and diluted 1:1000 for primary antibody incubation. Detailed information of commercial manufacturers and validation of the antibodies can be found in the Reporting Summary.