Figure 1.
Expression levels of ectipically expressed macroH2A1.1 and H2ABbd.
a, HeLa S3 cells overexpressing tagged macroH2A1.1 (1) as compared to endogenous macroH2A (2) in the control HeLa S3. Western Blot with staining with the anti- macroH2A antibodies (Active Motif); b, The expression of H2A.Bbd was measured using qRT-PCR in JKT1 human testicular seminoma cells, HeLa S3 and HeLaS3 ectopically expressing H2A.Bbd. Mean values are shown, error bars represent S.E.M. of 2 independent experiments.
Figure 2.
Subnuclear localisation of macroH2A and H2A.Bbd in HeLa S3cells.
HeLa S3 cells stably transfected with either the pOZ -macroH2A1.1 (a) or pOZ- H2A.Bbd (b) plasmid and stained with antibodies against FLAG (green) and H3k27Me3 or nucleophosmin (red). a: macroH2A1.1 is preferentially localised to a single region in interphase HeLa cells and coinsides with H3k27Me3 staining (red). b: H2A.Bbd has a punctate nuclear staining with an exclusion zone (marked by a white arrow) and a strong perinucleolar staining (nucleophosmin, red). The figure shows representative confocal sections. Scale bar: 5 µm.
Figure 3.
Distribution of sites of preferential location of macroH2A1.1 and H2A.Bbd within the telomeric end of the short arm of human chromosome 16.
(a) Low magnification picture showing the whole area under study. (b–f) High magnification (zoomed in) pictures showing consecutive sections of the area under study. Distribution of histone H3 tri-methylated at lysine 36 (H3K36me3), histone H3 tri-methylated at lysine 4 (H3K4me3), histone H3 di-methylated at lysine 4 (H3K4me2) and panacetylated histone H3 (H3ac) is presented according to the Pilot Encode datasets. Positions of genes are shown according to the UCSC genome browser (assembly HG18). For H3K36me3 and H3K4me3 two independent sets of data are shown. The values in brackets above the graphs showing distribution of macroH2A1.1 and H2A.Bbd represent the range of the transformed signal, (range of −log10(p-values) of the ACME p-values). The black broken lines in the sections showing distribution of macroH2A1.1 and H2A.Bbd indicate the significance level (−log10(p-value = 4)).
Figure 4.
Distribution of sites of preferential deposition of macroH2A1.1 and H2A.Bbd within the OR - β-globin gene domain in human chromosome 11.
(a) Low magnification picture showing the whole area under study. (b–f) High magnification (zoomed in) pictures showing consecutive sections of the area under study. All designations are the same as in Figure 3.
Table 1.
Presence of H2A.Bbd and macroH2A1.1 loading sites within gene bodies and promoter areas (2500 to +1000 and from −5000 to +1000) of genes present in the selected areas of the chromosome 11 and the chromosome 16.
Figure 5.
Distribution of H2A.Bbd and macroH2A1.1 on chromosomes 11 and 16.
H2A.Bbd (max(−log10(ACME-p-values)) (a) and macroH2A1.1 (max (−log10(ACME-p-values)) (b) in the gene body and short upstream area [−400; TSS] versus gene expression levels is shown for genes present in the selected areas of chromosomes 11 and 16. Figures to the right of gene positions show the number of significant peaks of macro H2A (a) and H2A.Bbd (b) present within the gene body (when this number was more than 1). See the text for details.
Figure 6.
Distribution of sites of preferential location of macroH2A1.1 and H2A.Bbd within the telomeric end of the chromosome 11 (a) and the region of the X chromosome containing the DMD gene (b).
The values in brackets above the graphs showing distribution of macroH2A1.1 and H2A.Bbd represent the range of the transformed signal, (range of −log10(p-values) of the ACME p-values). The black broken lines in the sections showing distribution of macroH2A1.1 and H2A.Bbd indicate the significance level (−log10(p-value = 4)).
Table 2.
The (H2A.Bbd/macroH2A1.1) ratio in 4 genomic regions studied.