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Supplementary Figure 3
Basal body localization of CPLANE proteins.
(a) Dorsal views of stage 19 Xenopus embryos show neural tube closure defects after Jbts17-knockdown that are rescued by expression of Jbst17-wild-type but not the Joubert-associated truncation (R1569*). The graph shows average distance between neural folds. (b) In situ hybridization to stage 30 Xenopus embryos shows loss of vax1, a gene downstream of sonic hedgehog, after Jbts17 knockdown; numbers in each panel indicate the number of embryos showing the phenotype. (c-f) Fluorescence micrographs show the localization of CPLANE proteins at basal bodies in Inturned (c), Fuzzy (d), Wdpcp (e) or Rsg1 (f) knockdown multi-ciliated cells. Green and red signals indicate GFP-tagged CPLANE proteins and centrin4-RFP, respectively. The graphs to the right in each figure show fluorescence intensity of GFP signals normalized against Centrin4-RFP (ns, non-significant; **P < 0.01; *** P < 0.001). (g) Fluorescence micrographs show the localization GFP-tagged Cep164, Ofd1, Hook2 and Mks1 in multi-ciliated cells in controls and after Jbts17 knockdown. The graph at right shows the normalized fluorescence intensity for the indicated proteins, as described for c-f. (h) CRISPR disruption of Jbts17 phenocopies MO knockdown. Gels at left demonstrate targeting of Jbts17 (see methods for details); images at right show disruption of ciliogenesis in MCCs.
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