SGOL1 average expression values obtained by RNA sequencing of three biological replicates after control and H2A.Z.2 siRNA treatment. Error bars show the standard deviation (SD). ns, not significant (Student's t‐test).
AURKB average expression values obtained by RNA sequencing of three biological replicates after control and H2A.Z.2 siRNA treatment. Error bars show the standard deviation (SD). ns, not significant (Student's t‐test).
Western blot analysis of Sgo1 and Aurora B in mitotic cells after control and H2A.Z.2 siRNA treatment.
CENP‐A average expression values obtained by RNA sequencing of three biological replicates after control and H2A.Z.2 siRNA treatment. Error bars show the standard deviation (SD). ns, not significant (Student's t‐test).
CENP‐C average expression values obtained by RNA sequencing of three biological replicates after control and H2A.Z.2 siRNA treatment. Error bars show the standard deviation (SD). ns, not significant (Student's t‐test).
Flow cytometry analyses profiles of control and H2A.Z.2 siRNA‐treated HeLa cells. Percentages represent the mean of two biological replicates. Data sets were statistically analysed using Chi‐square test. ns, not significant.
Mitotic index of HeLa cells transfected with control, H2A.Z.1 or H2A.Z.2 siRNA. Error bars represent SD of three biological replicates. At least 2,500 cells were analysed for each condition. Data sets were statistically analysed using Chi‐square test. ***P < 0.001; *P < 0.05.
Mitotic cells from the experiment in (G) were analysed and classified by mitotic stage. Error bars represent SD of three biological replicates. At least 300 mitotic cells were analysed for each condition. ns, not significant (Chi‐square test).
Mitotic spreads of HeLa cells treated with control (top) or H2A.Z.2 (bottom) siRNA. #1. The panel on the right shows a magnification of a chromosome from the box in the left panel (Scale bar 10 μm).
Violin plot of centromeric CENP‐C intensity of prometaphase/metaphase cells from HeLa cells treated with control, H2A.Z.2#1 or H2A.Z.2#2 siRNA (control si N = 243 H2A.Z.2 si#1 N = 80, H2A.Z.2 si#2 N = 502). The bars represent the median. Data sets were statistically analysed using the Wilcoxon rank test in R. ***P < 0.0001; ns, not significant.
Graph showing the correlation between YFP:CENP‐A signals and the intensity of Sgo1 signals at the centromeres of prometaphase chromosomes after H2A.Z.2 depletion. The black line represents the lowest smoothed fit.
Volcano plot representation of differentially expressed genes in the H2A.Z.1‐depleted cells vs the H2A.Z.2‐depleted cells data sets. Y‐axis represents the ‐log10 of the P‐value. Coloured points mark the genes with significantly increased or decreased expression. X‐axis represents the log2 value of the fold change: points with log2 < 0 indicate downregulated genes in the H2A.Z.2‐depleted cells data set compared with the H2A.Z.1‐depleted cells data set; log2 > 0 indicate upregulated genes (Student's t‐test).
Venn diagram of the regions identified by ATAC‐seq in control si, H2A.Z.1 si‐ and H2A.Z.2 si‐treated HeLa cells.
Representative image of HeLa nuclei after FISH with a Chr17 centromeric probe (green) (Scale bar 10 μm).
Quantification of the number of FISH signals/nucleus in control (grey) or H2A.Z.1‐depleted (blue) cells. At least 500 nuclei were analysed per condition. Data sets were statistically analysed using Fisher exact test. ns, not significant.
Frequency of genes with altered expression (upregulated—red and downregulated—green) per chromosome after H2A.Z.1 (top panel) or H2A.Z.2 (middle panel) depletion. Bottom panel shows the percentage of H2A.Z‐containing regions/chromosome. Data sets were statistically analysed using Fisher exact test. *P < 0.05, **P < 0.01, ***P < 0.001.
