Figure 2. H2A.Z.1ub is necessary for proper lineage commitment.

A. Bright field and H&E staining of day 12 embryoid bodies from H2A.Z.1^WT and H2A.Z.1^K3R3 mESCs. Arrows indicate examples of neuroectoderm present in H2A.Z.1^WT EB sections. B. IHC staining of d12 H2A.Z.1^WT and H2A.Z.1^K3R3 EB sections with Tuj1, a marker of neural differentiation. Arrows indicate examples of Tuj1 positive regions. Scale bar for all =100um. C. Expression of lineage markers during EB time course indicates lack of neuroectoderm differentiation in H2A.Z.1^K3R3 mESCs (red v. black lines). Addition of WNT inhibitor KY02111 to H2A.Z.1^K3R3 mESCs 48 hours prior to and during EB formation restores multi-lineage differentiation (yellow lines). Expression is analyzed by qRT-PCR with values normalized to H2A.Z.1^WT+DMSO using Tubb5. Error bars represent SD of triplicate reactions. See also Figure S3.