PRC1 and PRC2 proximal interactome in mouse embryonic stem cells - Western blots
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Raw images from all western blot analyses included in this manuscript
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Protein lysates were diluted in Laemlli sample buffer containing 100 mM DTT and bromophenol blue. Samples were boiled for 10 min at 95°C after which proteins were separated on an SDS–PAGE gel and transferred from the gel to a nitrocellulose membrane using a Trans-Blot Turbo Transfer System (Bio-Rad) according to the manufacturer’s instructions. Nitrocellulose membranes were blocked with 5% milk or BSA in 0.1% Tween-PBS for 1h followed by 1h primary antibody and 1h secondary antibody. Primary antibodies used were anti-EZH2 (Diagenode; C15410039), Strep-HRP (Invitrogen, S911), anti-HA clone 3f10 (Roche, 11867423001), anti-Tubulin (Sigma-Aldrich; T5168), anti-H4ac (Millipore; 06-598), anti-H3K27me3 (Diagenode; C15410195), anti-H3 (Abcam; ab1791), anti-GAPDH (Abcam; ab181602), anti-SUZ12 (Abcam; ab12073), anti-Actin (Abcam; ab8226), anti-RNF2 (Cell Signaling Technology; 70916), anti-H2AK119ub1 (Cell Signaling Technology; 8240), anti-SALL1 (Abcam; ab31526), anti-SALL4 (Abcam; ab29112), anti-NANOG (Cell Signaling Technology; 8785), anti-TRIM24 (Bethyl Laboratories; A300-815A).