(A) Western blots showing the levels of DNA methylation enzymes and regulators. Note that mESCs express two major Dnmt3a isoforms (Dnmt3a and Dnmt3a2) and that the Dnmt3a antibody cross-reacts with Dnmt3b.
(B) Nuclear fractionation assay showing Uhrf1 chromatin association. PCNA and HP1α were used as controls for soluble and chromatin-associated proteins, respectively.
(C) Quantification of data in (B) by densitometry using Image J. Shown are percentages of soluble and chromatin-associated Uhrf1 in each sample (mean + SD from three independent experiments).
(D and E) ChIP assays showing relative enrichment of H3R2me2a (D) and Uhrf1 (E) at MSR and IAP regions (mean + SD from three independent experiments).