Journal: EMBO Reports

Article Title: Expansion of the ISWI chromatin remodeler family with new active complexes

doi: 10.15252/embr.201744011

Figure Lengend Snippet: The expanded family of ISWI chromatin remodelers

Article Snippet: For each immunoprecipitation, ~8 × 10 6 cells were washed in phosphate‐buffered saline (PBS) and lysed for 30 min on ice in 50 mM Tris pH 7.5, 150 mM NaCl, 300 mM KCl, 10% glycerol, 1% Triton X‐100, freshly supplemented with 50 U/ml Benzonase (Sigma), 1 mM tris(2‐carboxyethyl)phosphine (TCEP), and protease inhibitors (cOmplete, EDTA‐free, Roche), with three 10 s vortex cycles every 10 min. Lysates were clarified by centrifugation at 14,000 × g for 15 min at 4°C and supplemented with 5 μg of antibody against SMARCA1 (#12483, Cell Signaling Technology), SMARCA5 (NB100‐55310, Novus Biologicals), BAZ1A (NB100‐61042, Novus Biologicals), BAZ1B (#2152, Cell Signaling Technology), BPTF (NB100‐41418, Novus Biologicals), or CECR2 (#730031, Thermo Fisher Scientific).

Techniques: