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Figure 3


Fig. 3. Gene knockdown of CK1{delta}/{epsilon} reduces the phosphorylation of Dvl2 induced by Wnt-5a. (A) SN4741 cells were transfected with control siRNA and three independent siRNAs against CK1{epsilon} isoform. The efficiency of gene knockdown induced by individual siRNAs against CK1{epsilon} was quantified in western blots. A non-specific protein, resulting in a band recognised by anti-CK1{epsilon} antibody served as a loading control. Quantification (normalised to loading control) is shown below. (B,C) SN4741 cells were transfected with the indicated combinations of siRNAs, and treated with control, 50 and 100 ng/ml of Wnt-5a. The phosphorylation of Dvl isoforms was detected as phosphorylation-dependent mobility shift of total Dvl2. Data are representative of three (B) or two (C) independent experiments. The position of dephosphorylated ({blacktriangleleft}) and phosphorylated Dvl ({triangleleft}) is indicated. The levels of CK1{epsilon} and beta-actin were also determined to confirm the efficient gene knockout and the equal protein loading.





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