Fig. 5

Phe/Ile/Asp amino acid residues of UBN1 and UBN2 are involved in the binding and deposition of H3.3. a Sanger sequencing of PCR product shows that Phe/Ile/Asp of both UBN1 and UBN2 were mutated to Alanine in FID-C4 cell line. b Venn diagram shows the overlapping among C4-H3.3, UBN-H3.3, and HIRA-H3.3. c, d Phe/Ile/Asp amino acids of UBN1/2 were important for the deposition of H3.3. Meta-analysis shows that alternation of the deposition of H3.3 at both promoters (c) and enhancers (d) in mES cells with FID/AAA mutation of UBN1 and UBN2. Reads were normalized to 10 million in each data set. (e, f) ChIP-qPCR analysis of H3.3 at enhancers and promoters of active (Nanog) (e) and bivalent (Hand1) (f) genes. The qPCR value was normalized to 1% input of each sample. Standard deviation was derived from three replicates