Down-regulation of DAXX, but not ATRX, abolishes recruitment of H3.3 to PML bodies. (A) Immunolocalization of DAXX and PML in cells expressing H3.3-mC, after sham (control) or siRNA-mediated DAXX knockdown. (B) Distribution of H3.3-mC fluorescence in a sham (control)– and DAXX-depleted cell (shown in inset, corresponding to H3.3-mCherry pictures in A). (C) Colocalization of coexpressed H3-3-EGFP and mC-DAXX with immunolabeled PML. (D) Immunolocalization of ATRX and PML in cells expressing H3.3-mC, after sham (control) or siRNA-mediated ATRX knockdown. (E) FRAP analysis of H3.3-EGFP in cells expressing mC-DAXX, in cells depleted of DAXX using two different siRNAs, and in sham-depleted cells (H2O) (mean ± SD of 10–14 cells). (F) FRAP analysis of H3.3-EGFP in ATRX-depleted (using two different siRNAs) and sham (H2O)–depleted cells, and in cells expressing mC-DAXX and depleted of ATRX (mean ± SD of seven cells). Scale bars, 10 μm.
