Fig. 3

Analysis of the effect of CKII phosphorylation upon DLG1 subcellular localization. A An intact E7 CKII phospho-acceptor site is required for DLG1 misdistribution in the presence of both HPV oncoproteins. Upper panel, the vector encoding egfp-DLG1 (1.5 μg) was transfected into HEK293 cells and DLG1 localization was analyzed by confocal microscopy (yellow arrow). Middle and bottom panel, the vector encoding mTurq2-18E6 (cyan 0.75 μg), egfp-DLG1 (green 0.75 μg) and wild type or mutant Cherry-18E7 (red 0.75 μg) were co-transfected into HEK293 cells and the localization of each fusion protein was analyzed by confocal microscopy after 24 post transfection, white and yellow arrows indicate DLG1 subcellular localization. B Effect of specific CKII inhibitor CX-4549 upon DLG1 localization. HEK293 cells were transfected with pegfp-DLG1 (green 1.5 μg), pmTurq2-18E6 (cyan 0.75 μg) and wild type pCherry-18 E7 (red 0.75 μg) for 24 hs. CX-4945 CKII inhibitor was added to the cells 4 h before fixing, as indicated (+ CX-4945). The bar graph shows the quantification of the DLG1 levels at cell borders in the presence or absence of the inhibitor CX-4945, at least 80 cells were analyzed per condition. All scale bars represent 10 μm. The results are representative of at least 3 independent experiments