Fig. 7
Wnt ligands promote open conformation of DVL3. a Western blot analysis of the effect of the Wnt-3a (3a) and Wnt-5a (5a) ligands on the phosphorylation of endogenous DVL2 and DVL3 in HEK293 wt cells. b, c Western blot analyses of samples from two stable cell lines (derived from HEK293 wt and HEK293 DVL1/2/3 triple knockout22 cells) that inducibly under tetracycline-controlled promoter express ECFP-DVL3 FlAsH III sensor. Dashed arrow indicates endogenous DVL3; full arrow indicates ECFP-DVL3. d Western blot analysis of the effect of the Wnt-3a (3a) and Wnt-5a (5a) ligands on the phosphorylation status of ECFP-DVL3 FlAsH III in HEK293 DVL1/2/3 triple knockout22 cells. e, f Measurements of the intramolecular FRET efficiency of the endogenously expressed DVL3 FlAsH sensors III after the treatment with Wnt ligands in HEK293 wt and HEK293 DVL1/2/3 triple knockout22 cells. One data point corresponds to one analyzed cell; datapoints from four independent transfections were merged. Data in e and f represent median ± interquartile range. Statistical significance in e and f was analyzed by one-way ANOVA test with Gaussian distribution and Tukey's post-test (*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001, ****, p ≤ 0.0001; ns, not significant, p > 0.05). CM conditioned medium, Inh. inhibitor, TC tetracycline
