Fig. 2: Effects of FZD5 micro-switch mutants on the electrophoretic mobility shift of DVL2. | Nature Communications

Fig. 2: Effects of FZD5 micro-switch mutants on the electrophoretic mobility shift of DVL2.

From: Pathway selectivity in Frizzleds is achieved by conserved micro-switches defining pathway-determining, active conformations

Fig. 2

a Immunoblots of HEK293A cells transiently transfected with the different FZD5 micro-switch mutants, wild-type FZD5, and pcDNA3.1 (Mock). Cell lysates were analyzed for the FZD5-induced electrophoretic mobility shift of DVL2 from basal (bottom band; filled triangle) to shifted (bracket, open triangle) using an anti-DVL2 antibody to detect endogenously expressed DVL2. Expression of full-length FZD5 constructs was detected using an anti−1D4 antibody. Anti-GAPDH served as a loading control. Note that the immunoblots are cropped. Wild-type FZD5 and Mock were included on each individual blot as a control. Uncropped blots are shown in Source Data File. b Quantification of basal and shifted DVL2 bands by densitometry. Data show mean ± SEM of six independent experiments for the ratio of phosphorylated and shifted (PS)/basal DVL2. Mean values were normalized to wild-type FZD5 on the respective immunoblot. Statistical differences between the micro-switch mutants and wild-type FZD5 were assessed by one-way ANOVA followed by Dunnett’s post-hoc analysis. Significance levels are given as follows: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. TM transmembrane domain, ECL extracellular loop, H8 helix 8.

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