Fig. 7.

Both USP15 isoforms are predominantly cytosolic but localise to the nucleus at prophase. A549 cells were transfected with the indicated GFP-USP15 constructs or GFP only as a control. Cells were fixed and co-stained for the cell cycle phase marker cyclin B1 and DAPI, to classify as G1 or S-phase (low cytoplasmic cyclin B1), G2 (high cytoplasmic cyclin B1), or prophase (nuclear cyclin B1, uncondensed chromatin). a Representative immunofluorescence images of cells expressing the two wild-type (WT) USP15 isoforms in G2 cells and prophase cells; scale bars 10 μm. b, c Transfected cells were scored for relative intensity of GFP-USP15 variant expression in the nucleus compared with the cytoplasm, for cells in G2 b or prophase c; counts for >100 cells per GFP-USP15 construct were pooled from three independent experiments, mean and SD are indicated. Additional images are shown in Supplementary Figure S4