Hat1 is required for the acetylation of histones H3 and H4 deposited during replication-coupled chromatin assembly.

<p>A and B) Hat1^+/+ and Hat1^−/− MEFs were pulse-labeled with EdU and chased with thymidine as schematically depicted at the top of each panel. Samples were isolated at the indicated time points and proteins associated with nascent DNA were resolved by SDS-PAGE following affinity purification of EdU-labeled DNA (iPond). The indicated amounts of the input fractions (prior to affinity purification) are on the left-hand side of each panel. Samples eluted from the affinity purification resin are on the right-hand side of each panel. In all cases, the respective Hat1^+/+ and Hat1^−/− samples were analyzed on the same gel (separated by a MW standard lane that has been removed). Western blots were probed with the antibodies indicated on the right. Numbers below each lane indicate the normalized intensity of the band as determined using Licor software. Each sample was normalized to the level of unmodified histone H3. The intensity observed in the 30′ pulse sample was arbitrarily set to 1.0. No Clck indicates control samples that were not biotin labeled.