Anti wdr5

For nuclear lysate preparation, cells were harvested, washed in PBS, and resuspended in Buffer A (10 mM HEPES (pH 7.9), 10 mM KCl, 1.5 mM MgCl₂, 0.34 M sucrose, 10% glycerol and 0.1% Triton) for 8 min on ice. Nuclei were centrifuged at 1300 × g and resuspended again in Buffer A containing benzonase. To prepare whole cell lysates, cells were harvested, washed and re-suspended in RIPA buffer (50 mM Tris (pH 8), 150 mM NaCl, 0.1% SDS, 1 mM EDTA, 0.5% DOC and 1% NP-40) for 30 min on ice. Nuclei were centrifuged at 12 000 RPM, and the supernatant was collected. Protein concentration was determined using the Bradford Assay (Bio-Rad). All capillary electrophoresis runs were normalized to Lamin (anti-Lamin, Cell Signaling 2032) or total protein (Protein Simple). Primary antibodies used include: anti-Treslin (Bethyl A303-472A), anti-MTBP (Santa Cruz, sc-137201), anti-GFP (Abcam, ab290), anti-CUL4B (Sigma, HPA011880), anti-CUL4A (Cell Signaling, 2699), anti-DDB1 (Santa Cruz Biotechnology, SC-137132), anti-CDT2 (Abcam, ab184548), anti-PCNA (Santa Cruz Biotechnology, PC10, sc-56), anti-CDC45L (Protein Tech 15678-1-AP), anti-WDR5 (Cell Signaling, 13105), anti-CDT1 (Abcam, ab202067), anti-p21 (Cell Signaling, 2947) and anti-SET8 (Cell Signaling, 2996). Images were generated with Compass Software (Protein Simple).