Fig. 2

Phosphorylation of 18E7 by CKII is required for stabilization of DLG1 levels. HEK293 cells were transfected with 1 μg of pmTurq2-DLG1, 5 μg of pseyfp-18E6 and/or 5 μg of wild type or mutant pCMV-Flag-18 E7, as indicated. Fusion proteins were assessed by western blot 24 hs post transfection (Upper panel) using an anti-Flag (E7), anti-18E6 and anti-GFP, which recognizes mTurq2-DLG1. β-Galactoside levels were used as transfection control. When indicated, CX-4945 CKII inhibitor was added to the cells 4 h before harvesting. Images with different exposure levels area shown in Fig. S5. Bottom panel, densitometry analysis of DLG1 levels normalized to β-Galactoside expression. Asterisks denote the significant difference between conditions tested by ANOVA test and a Multiple Comparison Tukey´s test (***p < 0.005, ****p < 0.0005) and “ns” indicates no significant changes. The results are representative of three independent experiments. Original blots are presented in Fig. S5