Figure 5.

Changes in Levels of H4ac and H3K4met Are Limited to Specific Loci in HDA6 Mutants.

(A) Protein gel blot analysis detecting H4ac (top panel) and H3K4met (middle panel) using α-H4ac and α-H3K4met antibodies, respectively, on protein extracts from wild-type (DR5) and axe1-5 mutant plants. Bottom panel, Coomassie staining shows equal protein loading.

(B) ChIP performed in the control line DR5 and the mutant allele axe1-5 reveals an increase in H4ac and H3K4met at rDNA repeats. The Actin2/7 gene is equally present in mutant and control precipitates. If the antibodies are omitted during the procedure (mock), neither target is amplified, whereas the equal strength of bands after PCR with the input fraction indicates equal amounts of chromatin before immunoprecipitation.

(C) and (D) Distribution of histone H3 methylated at Lys 4 revealed by DAPI staining (blue, left panel) and immunodetection with an antibody specific for H3K4met (green, middle panel) in nuclei of control lines (top row) DR5 (C), Ler (D), and mutants (bottom row) axe1-5 (C) and sil1 (D). Right panels show merged images.