Figure 7

ChIP analyses of permissive histone modifications in macronuclear anlagen. (A) Diagram of the micronuclear and the macronuclear version of the Stylonychia TEBPα gene. MDSs occur in scrambled disorder (1-3-5-7-9-11-2-4-6-8-10-12-13-14) in the micronucleus. For real-time PCR analyses primers from MDS3 (Mi2) and IES3 (IES3) were used. (B) Diagram of the micronuclear and the macronuclear version of the Stylonychia 1.1 kb and 1.3 kb locus. For real-time PCR analyses of the MDSs primers from the 1.1 kb (P94) and the 1.3 kb (P95) locus were selected. For real-time PCR analyses of the flanking sequence primers Sz1 and Sz2 were used. (C) Diagram of the micronuclear and the macronuclear version of the Stylonychia actin I gene. MDSs occur in scrambled disorder (3-4-5-6-7-8-10- [-2]- [-1]-9) in the micronucleus, furthermore MDSs 1 and 2 are inverted. For real-time PCR analyses of the MDS primers from MDS10 (Actinfor) and MDS-2 (Actinrev) were used. For real-time PCR analyses of the flanking sequence primers Actin5f and Actin5r were selected. (D) Diagram of the micronucleus specific sequence stad5. For real-time PCR analysis primers Stad5-3654 and Stad5-3520 were used. (A-D) Positions of PCR fragments are marked above the diagrams. (E) Fluorescence in situ hybridization on isolated nuclei using stad5 as a probe. Stad5 (green) could be detected in micronuclei (m) and macronuclear anlagen (stage a₂), but not in macronuclei (M). (F) Quantitative real-time PCR analyses of ChIP experiments. Primer sequences are listed in Methods.