FIGURE 6.

NES deletion in monomeric PCNA NES is functional in monomeric PCNA. HeLa cells were transfected with pcDNA3-GFP-wtPCNA (WT PCNA) or pcDNA3-GFP-PCNAY114A (PCNAY114A). A, effect of LMB on the monomeric PCNA mutant PCNAY114A. HeLa cells expressing PCNAY114A were treated with either ethanol solvent (−) or 20 ng/ml LMB for 6 h prior to fixation and permeabilization, and cells were analyzed by scanning confocal microscopy. HeLa cells transfected with pcDNA3-GFP-wtPCNA and treated with solvent were used as controls. B, colocalization of CRM1 with PCNAY114A but not with WT PCNA in HeLa cells. Immunofluorescence detection of CRM1 was performed using a rabbit polyclonal anti-CRM1 antibody followed by an Alexa Fluor 555-coupled goat anti-rabbit IgG, and cells were analyzed by scanning confocal microscopy. A and B show representative experiments that have been performed at least three times with similar results. The nuclei were visualized with Hoechst staining (original magnification ×630, scale bar = 20 μm).