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Extended Data Fig. 8 | H2A.Z is essential for DNA replication and cell
proliferation during T-cell activation. a, Diagram showing the construction
of mice with H2A.Z conditionally knocked out in T cells. b, FACS analysis of
T cells and B cells from the spleen of wild-type (H2A.Zf/f) or H2A.Z CKO
(CD 4creH2A.Zf/f) mice. c, Statistical analysis of T-cell numbers from panel b.
d, Left, FACS analysis of BrdU incorporation in CD8+ T cells from the lymph
node of wild-type and H2A.Z CKO mixed bone-marrow chimaeric mice. Right,
statistical analysis of the percentage of BrdU-incorporating cells. FSC, forward
s c a t t e r. e, FACS analysis of CFSE dilution in wild-type and H2A.Z CKO CD4+
T cells upon anti-CD3 and anti-CD28 stimulation for 72 h. f, Statistical analysis
of the percentage of divided cells from panel e. g, Cell-cycle analysis of wild-
type and H2A.Z CKO CD4+ T cells upon anti-CD3 and anti-CD28 stimulation for
48 h. PI, propidium iodide. h, Statistical analysis of cell-cycle distribution from
panel g. i, Heat map and box plots showing the H4K20me2 and NS signals in 10-
kb regions around H4K20me2 peaks (n = 53,788) and NS peaks (n = 26, 2 83). j,
Western blots showing H4K20me2 levels in active T cells from wild-type and
H2A.Z CKO mice. k, Box plots showing H2A.Z, H4K20me2 and NS signals in
ZD-NS (n = 18,382) and ZI-NS (n = 7,901) regions. Data in panels c, d, f and h are
means ± s.e.m.; n = 4 biological replicates in panel c; n = 5 biological replicates
in panel d; n = 3 biological replicates in panels f, h; two-tailed unpaired t-test.
The data in panels i, k were analysed by two-tailed paired t-test and two-tailed
unpaired t-test, respectively. FACS analyses in panels b, d, e, g were
independently repeated four, five, three and three times, respectively, with
similar results. The western blots in panel j were independently repeated three
times with similar results, and H4 was used as a loading control and sample
processing control. For the box plots in panels i, k, the centre lines represent
medians, the box limits are the 25th and 75th percentiles, and the whiskers are
the minimum to maximum values. For gel source data, see Supplementary
Fig. 1. For FACS gating strategy, see Supplementary Fig. 3.