Primary antibodies used were: mouse monoclonal anti-PAR (Trevigen,4335-MC-100), mouse monoclonal anti-Myc (9E10 clone, hybridoma-conditioned medium), rat monoclonal anti-α tubulin (Santa Cruz Biotechnology, sc-53030), rabbit polyclonal anti-acetyl-histone H3, recognizing histone H3 acetylated at the N-terminus (Millipore, 06–599), rabbit polyclonal anti-acetyl-histone H4, recognizing all 4 lysine residues (Millipore, 06–866), rabbit polyclonal anti-histone H3, recognizing the C-terminal region of histone H3, (Millipore, 07–690), rabbit polyclonal anti-histone H4, recognizing the C-terminal region of histone H4 (Abcam, ab10158). Secondary antibodies used were: goat anti-mouse, anti-rabbit and anti-rat horseradish peroxidase-conjugated (Santa Cruz Biotechnology).
Detection was carried out using the ECL Western blotting detection reagents (GE Healthcare). Densitometric analysis was performed using Image Lab software (Bio-Rad). For each lane, the value corresponding to the area of histone H3 or H4 band in the membrane probed with anti-acetyl-histone H3 or H4 was normalized by dividing for the value corresponding to the area of histone H3 or H4 band in the same membrane re-probed, after mild stripping, with anti-C-terminal region of histone H3 or H4.
Detection was carried out using the ECL Western blotting detection reagents (GE Healthcare). Densitometric analysis was performed using Image Lab software (Bio-Rad). For each lane, the value corresponding to the area of histone H3 or H4 band in the membrane probed with anti-acetyl-histone H3 or H4 was normalized by dividing for the value corresponding to the area of histone H3 or H4 band in the same membrane re-probed, after mild stripping, with anti-C-terminal region of histone H3 or H4.