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. 2011 Jan 31;6(1):e16598. doi: 10.1371/journal.pone.0016598

Figure 1. USP7 regulates PML-NBs independent of its ubiquitin cleavage activity.

Figure 1

(A) CNE2 cells were treated with siRNA against USP7 (siUSP7) or GFP (siGFP) then cells were fixed and stained for PML and endogenous USP7. The number of PML-NBs was counted for a minimum of 50 cells for each sample in three independent experiments. Histogram on the right represents the average distribution of number of PML-NBs in siGFP (white bars) and siUSP7 (black bars) samples, where error bars represent standard deviation from the three independent experiments. Images shown are for siUSP7 samples in which green stained cells serves as an internal control for cells in which USP7 expression has not been silenced. (B) CNE2 cells were transiently transfected with 2 µg of a plasmid expressing WT USP7 or USP7 domains as indicated or with the empty plasmid (Mock). 24 hours post-transfection, cells were fixed and stained using PML (red) and c-Myc (green) antibodies. Effect of USP7 overexpression on the number of PML-NBs was quantified as in (A), except at least 100 cells were counted for each sample.