Mabc1152

Cells were pelleted by centrifugation at 228g for 5 min at 4 °C and resuspended in RIPA buffer (150 mM sodium chloride, 1.0% NP-40, or Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS (sodium dodecyl sulfate) and 50 mM Tris, pH 8.0) and protease inhibitors with benzonase (Novagen; final concentration, 1.25 U µl^–1) and incubated for 30 min on ice with intermittent mixing. Extracts were sonicated for 5 cycles with Bioruptor (Diagenode) with the 30 s on and 30 s off cycles, and were cleared by centrifugation at 15,500g for 10 min at 4 °C. Equal amounts of protein extract were denatured in 1× Bolt LDS sample buffer (Thermo Fisher Scientific, B0007) and separated on Bolt Bis-Tris gels (Thermo Fisher Scientific, NW04120BOX, NW00122BOX), blotted on a polyvinylidene fluoride (PVDF) membrane (BioRad, 1704156) and immunoblotted with antibodies to MSL3 (Merck Millipore, ABE467, 1:1,000 dilution), L1 ORF1 (Merck Millipore, MABC1152, 1:1,000 dilution), H4K16ac (Abcam, ab109463, 1:5,000 dilution), H3K27ac (Abcam, ab4729, 1:5,000 dilution), ɑ-tubulin (Sigma, T9026, 1:5,000 dilution), and HERV (Novus Biologicals, NB100-93579, 1:500 dilution), and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG H&L (Abcam, ab6721) and HRP-conjugated goat anti-mouse H&L (Thermo Fisher Scientific, 31430) secondary antibodies.