Disrupted ciliogenesis caused by loss of zcrb1 is independent of Wnt signaling up-regulation in zebrafish.
(A) Quantification of gastrulation and body axis phenotypes in zcrb1 crispant versus Cas12a control–injected embryos in the presence of the WNT inhibitor XAV-939 or DMSO vehicle control. n > 20 embryos per condition. A minimum of 3 independent rounds of injection were analyzed (scale bar: 500 μm). (B, C). Representative images (B) and quantification (C) of Wnt signaling activation using the Tg(7xTCF-Xla.Sia:NLS-mCherry)ia5Tg transgenic zebrafish line assessing activation in Cas12a-injected embryos, zcrb1 crispant embryos, embryos injected with ZCRB1 WT human mRNA, and zcrb1 crispant embryos co-injected with ZCRB1 WT human mRNA. The quantification represents the mean fluorescence intensity of the reporter line (arbitrary units). P-values were measured by an ordinary one-way ANOVA with Tukey’s multiple comparisons test. Dots represent individual embryos per condition. A minimum of two independent rounds of injection were analyzed (scale bar: 500 μm). (D, E) Representative images (D) and quantification (E) of the percent number of primary cilia in zcrb1 crispant versus Cas12a control–injected embryos in the presence of the WNT inhibitor XAV-939 or DMSO vehicle control. Graphs are box plots with min/max error bars. P-values were measured by an ordinary one-way ANOVA with Tukey’s multiple comparisons test (scale bar: 100 μm).
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