Skip to main content
. 2021 Aug 23;22(11):e52061. doi: 10.15252/embr.202052061

Figure 1. H2A.Z.2.1 knockdown leads to genome instability.

Figure 1

  1. Representative images of HeLa cells treated with control, H2A.Z.1 or H2A.Z.2 siRNA for 72 h, fixed and stained with DAPI. White arrows point at micronuclei. Scale bar: 10 μm.
  2. Quantification of the percentage of cells with micronuclei from experiment in (A). Three biological replicates were analysed for each condition (control si N = 1,271; H2A.Z.1 si N = 1,091; H2A.Z.2#1 si N = 1,188; H2A.Z.2#2 si N = 1,193). The error bars represent the SD. Data sets were statistically analysed using Chi‐square test. ***P < 0.001.
  3. Quantification of the percentage of cells with micronuclei from HeLa cells co‐transfected with H2A.Z.2 siRNA and GFP:H2A.Z.2.1WT with either single or double mutations. Error bars represent the SD of three biological replicates (control si N = 1,316; H2A.Z.2 si N = 1,877; H2A.Z.2 si + wt N = 1,973; H2A.Z.2 si + A14T N = 618; H2A.Z.2 si + S38T N = 558; H2A.Z.2 si + A127V N = 649; H2A.Z.2 si + A14T/S38T N = 1,090; H2A.Z.2 si + A14T/A127V N = 1,038; H2A.Z.2 si + S38T/A127V N = 1,150). Data sets were statistically analysed using Chi‐square test. ***P < 0.001; black stars refer to the comparison with the H2A.Z.2si data, and blue stars refer to the comparison with the H2A.Z.2si + wt data; no stars = non‐significant.
  4. Schemes of the GFP constructs used for the rescue experiments in (E) and (F). Green boxes represent the GFP, brown boxes represent the H2A.Z.2.1 isoform. Solid fill represents the WT construct whereas striped box represents the KR mutant form where the residues indicate the mutations performed. Dots represent the possible post‐translational modifications in the mutated amino acids: acetylation (ac, dark blue), methylation (me, pink) and ubiquitination (ub, light blue).
  5. Representative images of prometaphase chromosomes from HeLa cells co‐transfected with H2A.Z.2 siRNA and each of the constructs in (D) (green). Scale bar: 5 μm.
  6. Quantification of the percentage of cells with micronuclei from experiment (E). The error bars represent the SD of three biological replicates (control si N = 887; H2A.Z.2 si N = 1,005; H2A.Z.2 si + wt N = 760; H2A.Z.2 si + KR N = 479). Data sets were statistically analysed using Chi‐square test. ***P < 0.001; ns, not significant. Blue stars refer to the comparison with the H2A.Z.2 si + wt data.