Nuclear localization of H3 variants and post-translational modifications (PTMs) and results of chromatin immunoprecipitatino (ChIP) analyses on chromatin purified from polytene anlagen (a3) using a pAb targeted to H3K36ac. (A) Western blot analyses of nuclear proteins isolated from micronuclei (m), macronuclei (M) and polytene anlagen (a3). Antibodies used were targeted against H3.3, H3.5, H3.7, H3K4me3, H3K9ac/K14ac, H3K27me3, H3K36ac, and H3S10ph. Red arrows indicate bands corresponding to 20 kDa or 15 kDa H3 variants. (B) Results of Western blot analyses post-immunoprecipitation. Antibodies targeted to H3K36ac were used for ChIP on polytene anlagen (a3) chromatin. Immunocomplexes were separated by SDS-PAGE and analyzed by Western blots using anti-H3.7 pAbs for detection. Left lane: protein size standard visualized by direct light photography; right lane: fluorescence light detection of a 20 kDa protein band. (C) Enrichment of several model sequences representing MDSs (H4, ACT1, two loci within MDP2) or micronucleus-specific sequences (MaA81, Stad5, pLJ01) was studied. Box plots exhibit median, interquartile range, and minimum and maximum values.