ChIP-seq from two biological replicates of control and Lam-KD S2 cells with anti-H3-pan acetylated antibodies (Active Motif, #39139) was performed as previously described (Chanas et al., 2014), with some modifications. After rinsing with PBS, ~2×107 cells were fixed with 1.8% formaldehyde in PBS containing 0.5 mM DTT for 20 min at room temperature. Cross-linking was stopped by adding glycine to 225 mM for 5 min and washing in PBS containing 0.5 mM DTT three times for 5 min. Cells were washed once in the A2 buffer (140 mM NaCl, 15 mM HEPES pH 7.6, 1 mM EDTA, 0.5 mM EGTA, 1% Triton X-100, 0.1% sodium deoxycholate, 0.5 mM DTT, complete EDTA-free protease inhibitor cocktail (Roche)). Cells were lysed in the A2 buffer containing 1% SDS for 10 min at room temperature, after that the lysate was 20-fold diluted by the A2 buffer and incubated for 2 min at 4°C. After sonication with VCX 400 Vibra-Cell Processor (Sonics; 30 pulses of 10 sec with 10-sec intervals at 15% max power) and 10-min high-speed centrifugation, the fragmented chromatin (with the average DNA fragment size ~0.5 kb) was recovered in the supernatant. For each immunoprecipitation, ~10 μg of chromatin (~700 µl) was pre-incubated in the presence of 100 μl of Protein A-Sepharose (PAS, 50% w/v, GE Healthcare) for 1 h at 4°C. PAS was removed by centrifugation, 5% of chromatin was isolated as an “Input” material, after that 2 µl anti-H3-pan acetylated antibodies (Active Motif, #39139) were added to the rest chromatin and samples were incubated overnight at 4°C in a rotating wheel. Then, 100 μl of PAS was added and incubation was continued for 4 h at 4°C. Samples were centrifuged at maximum speed for 1 min and the supernatant was discarded. Samples were washed four times in the A2 buffer containing 0.05% SDS and twice in 1 mM EDTA, 10 mM Tris (pH 8), 0.5 mM DTT buffer (each wash for 5 min at 4°C). Chromatin was eluted from PAS in 100 μl of 10 mM EDTA, 1% SDS, 50 mM Tris (pH 8) at 65°C for 10 min, followed by centrifugation and recovery of the supernatant. PAS material was re-extracted in 150 μl of TE, 0.67% SDS. To reverse cross-links, the combined eluate (250 μl) was incubated 6 h at 65°C and treated by Proteinase K for 3 h at 50°C. Samples were phenol-chloroform extracted and isopropanol precipitated in the presence of 20 μg glycogen. DNA was dissolved in 100 μl of water. ChIP samples containing ~25 ng of precipitated DNA, as well as “Input” samples were prepared for next-generation sequencing using a NEBNext Ultra II DNA library prep kit for Illumina (New England Biolabs).