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Fig. 4 | Epigenetics & Chromatin

Fig. 4

From: Histone H3K9 and H4 Acetylations and Transcription Facilitate the Initial CENP-AHCP−3 Deposition and De Novo Centromere Establishment in Caenorhabditis elegans Artificial Chromosomes

Fig. 4

Histone deacetylase HDA-1 tethering reduces RNA polymerase II-mediated transcription initiation signal on newly formed ACs. Immunofluorescence of RNA polymerase II Serine 5 phosphorylation (RNA Pol II Ser5P) on first-generation ACs at different cell stages, and ACs that have been propagated for generations and endogenous chromosomes in GFP::LacI-, GFP::LacI::HDA-1- and GFP::LacI::HDA-1(H145A)-tethering strain. Cropped images containing ACs and endogenous chromosomes (Endo Chr.) were shown. Embryos were stained with antibody against RNA Pol II Ser5P (red), antibody against LacI (green) and DAPI (blue), shown separately and merged. Scale bar represents 1 μm for both ACs and endogenous chromosomes. Quantification of IF signals. RNA Pol II Ser5P signals were normalized with DAPI signals, and the mean normalized RNA Pol II Ser5P signal intensity was calculated. The number of cells (n) analyzed was indicated. Error bars indicate 95% confidence interval (CI) for the mean. ***p < 0.001, **p < 0.01 and *p < 0.05 by Student’s t test. NS means not significant. Black arcs show comparisons between GFP::LacI- and GFP::LacI::HDA-1-tethering strains at the same cell stage. Blue arcs show comparisons between ACs at different stages in GFP::LacI-tethering strain

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Epigenetics & Chromatin

ISSN: 1756-8935