Figure 4: A. Ddx5 and Ddx5MUT both interact with Dnmt3a2 in vitro. Recruitment assay performed with the yeast Gal4 (DBD) and Gal4-Dnmt3a2 recombinant proteins bound on a biotinylated DNA fragment containing five tandemly repeated Gal4 binding sites, which was immobilized on streptavidine coated magnetic beads. His-Ddx5 (lanes 4-6 first panel) and His-Ddx5MUT (lanes 4-6 second panel) proteins are recruited to GAL4-DBD-tagged Dnmt3a2 protein-loaded beads but not to the control GAL4- DBD loaded beads (lanes 1-3 first and second panel respectively). His-tagged protein recruitment was detected with the α-Xpress antibody. α-Gal4 antibody indicates the input amounts of Gal4-DBD (lanes 1-3, third panel) and Gal4- Dnmt3a2 proteins (lanes 4-6, third panel) in the first and second panel samples. B. RFP-tagged Ddx5 and Ddx5MUT colocalize with GFP-tagged Dnmt3a1 and Dnmt3a2 proteins. First and third panels: Confocal microscopy of EGFP-Dnmt3a1 together with RFP-Ddx5 and RFP-Ddx5MUT proteins. Second and fourth panels: Confocal microscopy of EGFP-Dnmt3a2 together with RFP-Ddx5 and RFP-Ddx5MUT proteins.