For IP, whole-cell lysates were extracted using EBC lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 0.5% NP-40; freshly supplemented with protease and phosphatase inhibitor mixtures) for 30 min at 4°C, followed by centrifugation (12000 g for 10 min). For IP analysis of exogenous protein-protein interactions, whole-cell lysates extracted from stable cells transfected with Flag-tagged PAF or RBBP4 were incubated for 2 h with 15 μL of M2 magnetic beads (Sigma; M8823). For IP analysis of endogenous protein-protein interactions, whole-cell lysates extracted from control or stably transfected KP or A549 cells (shGFP or shPAF) were incubated overnight with protein G Dynabeads (Thermo) and 2∼5 μg antibodies against PAF (Santa Cruz), PAF (Abcam), RBBP4 (GeneTex), RBBP4 (Bethyl), p130 (Cell Signaling), E2F4 (Bethyl), LIN54 (Bethyl), BMYB (Abcam), and FOXM1 (Santa Cruz). After three to five washes with EBC lysis buffer, precipitates were eluted with SDS sample buffer and analyzed using immunoblotting. For analysis of PAF protein levels in NSCLC and PAF KD cells, radioimmunoprecipitation assay buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, and protease and phosphatase inhibitor mixtures) was used for whole-cell lysate preparation. For in vitro transcription and translation and binding assays, FLAG-PAF, FLAG-RBBP4, and HA-p130 were obtained with an in vitro TnT coupled system (Promega) and immunoprecipitated using anti-HA magnetic beads (Thermo). Immunoblot blocking and antibody incubation were conducted using 2% bovine serum albumin or 5% nonfat dry milk in TBST (25 mM, pH 8.0, 125 mM NaCl, and 0.1% Tween-20). SuperSignal West Pico and Femto (Thermo) were used to detect horseradish peroxidase-conjugated secondary antibodies. The following antibodies were used for immunoblotting: PAF (Santa Cruz, 1:5000), RBBP4 (GeneTex, 1:10000), p130 (Cell Signaling, 1:5000), E2F4 (Bethyl, 1:10000), E2F4 (Santa Cruz, 1:2000), LIN9 (Santa Cruz, 1:2000), LIN54 (Bethyl, 1:10000), FLAG M2 (Thermo,1:20000), and HA (Roche, 1:2000).

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