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Fig. 3. Suppression of DVL1 and DVL3, but not of DVL2, abolishes the ability of WNT3a to stimulate JNK activation, whereas expression of exogenous human DVL1 and DVL3 rescues the effect. F9 cells expressing rFZ1 were treated with siRNAs designed to suppress the expression of DVL1 (A), DVL2 (B) and DVL3 (C) for 48 hours, and JNK activity was measured by probing Jun phosphorylation, as described in the Materials and Methods. Specific suppression of individual DVL isoforms was demonstrated by immunoblotting with isoform-specific antibodies. The extent of suppression of DVL isoforms is as follows; DVL1 (71%), DVL2 (81%) and DVL3 (85%). The siRNAs were specific to the particular isoform as no cross-reaction was detected (D). Rescue experiments were performed by transfection of human (h) DVL1 (E) or DVL3 (F) into rFZ1-expressing cells in which DVL1 and DVL3 were knocked-down, respectively, by siRNA treatment as described in the Materials and Methods. The upper panel displays mean values±s.e.m. obtained from three independent experiments; the lower panel displays the corresponding representative blots.*P<0.05; **P<0.01 versus –WNT3a control; #, P<0.05; ##, P<0.01 versus +WNT3a control.
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