Figure 2. PPM1D overexpression induces an increase in nucleolar number.
H1299(PMD-9) and H1299(PMD-12) were derived from H1299 in our laboratory and expressed HA-PPM1D stably. H1299 cells were solubilized with 1x sample buffer for Western blotting (a) or fixed with formaldehyde for Immunocytochemistry (b). (a) PPM1D proteins were analysed by using Western blotting with rabbit polyclonal anti-PPM1D. (b) The subcellular localizations of NPM and PPM1D. H1299 cells and clones were fixed and stained with rabbit polyclonal anti-PPM1D and mouse monoclonal anti-NPM. (c) The percentage of cells with indicated nucleoli in H1299 clones were shown. For the quantitative analysis of the nucleolar number, stained-NPM was used as an indicator of the nucleoli. Number of cells used in the analysis are shown in Table 2; from at least two independent experiments.