Supplemental Figure 1
-
Fig. S1. Interaction between Dnmt3a and Dnmt3b. (A) Total
cell extract from J1 ES cells (middle panel) or Dnmt3b-/-
ES cells (8bb; right panel) was immunoprecipitated using an anti-Dnmt3b
antibody, and the immunoprecipitates were immunoblotted with an anti-Dnmt3a
antibody, which also cross-reacts with Dnmt3b. The left panel shows the results
of immunoblotting in which the total cell extract from J1 ES cells and 8bb
cells was analyzed using the anti-Dnmt3a antibody. (B) Schematic diagrams of mouse Dnmt3a, Dnmt3b, and
the mutant proteins we generated. The conserved PWWP and ATRX domains, and the
methyltransferase motifs (I, IV, VI, IX and X) are indicated. The location of
the Dnmt3a-Dnmt3b interaction domain determined from C is also shown. (C)
Interaction between Dnmt3a and Dnmt3b. COS-7 cells were transfected with two
expression vectors, one for myc-Dnmt3a or myc-Dnmt3b1, and another for
GFP-Dnmt3a, GFP-Dnmt3b1 or GFP-fused Dnmt3a/3b mutants, as indicated.
GFP-tagged proteins were immunoprecipitated from cell extracts using an
anti-GFP antibody. Immunoblotting analysis of the immunoprecipitates was
carried out using anti-myc antibody (top panel). The middle and bottom panels
show the results of immunoblotting of the total cell extract from transfected
cells, using anti-myc and anti-GFP antibodies, respectively.
