Figure 2.

De Novo Accumulation of H2A Variants at Replication Foci

(A–D) New histone accumulation at replication foci marked by CAF-1 p150 analyzed in U2OS cells stably expressing SNAP-tagged H2A.X, wild-type form (A–C) or different H2A variants (D) and treated with the indicated RNAPII inhibitors (DRB, flavopriridol [FLV], α-amanitin [AMA], and DMSO and PBS [vehicles]). Replication patterns (B) distinguish early, mid-, and late S phase cells. Total H2A.X is detected by immunostaining for SNAP or H2A.X (C). Bar chart, mean ± SD from 2 independent experiments. Scale bars, 10 μm. ns, non-significant; ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001. See also Figures S1–S3.