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Fig. 3
Dchs1b Interacts with the N-Terminal TPR Motifs of Ttc28 via its ICD CM2-N Motif
(A) RT-PCR assays to compare the expression levels of ttc28 at different developmental stages in zebrafish. gapdh housekeeping gene as a reference control.
(B) The expression patterns of ttc28 revealed by whole-mount in situ hybridization at eight-cell stage (1.15 hpf), 4.3 hpf, and 1 dpf zebrafish. ttc28 sense probe was used as a negative control.
(C) Co-IP assays of HA-Ttc28 with full-length Dchs1b-sfGFP or membrane GFP.
(D) Co-IP assays of Flag-Dchs1b-ICD with HA-Ttc28.
(E) Co-IP assays of Flag-Dchs1b-ICD with HA-Ttc28-N, HA-Ttc28-N-1, or HA-Ttc28-ΔN-1.
(F) Schematic of Ttc28 truncation constructs and the co-IP results of Ttc28 deletion mapping.
(G) Co-IP assays of HA-Ttc28-N with Flag-Dchs1b-ICDΔCM1, Flag-Dchs1b-ICDΔCM2, or Flag-Dchs1b-ICDΔCM3.
(H) Co-IP assays of HA-Ttc28-N with Flag-Dchs1b-ICD-N1, Flag-Dchs1b-ICD-N2, or Flag-Dchs1b-ICD-N3.
(I) Co-IP assays of HA-Ttc28-N with Flag-Dchs1b-ICD, Flag-Dchs1b-ICDΔCM2-N, or Flag-Dchs1b-ICDΔCM2-C.
(J) Schematic of Dchs1b-ICD deletion and truncation constructs and sequence alignment of Dchs CM2 motifs from different species. Disorder consensus bar, intensity of green corresponds with the strength of the consensus disorder agreement among eight prediction algorithms (Oates et al., 2013). Red line, residues of zebrafish Dchs1b involved in Ttc28 interaction.
See also Figures S2 and S3.
Reprinted from Developmental Cell, 45, Chen, J., Castelvecchi, G.D., Li-Villarreal, N., Raught, B., Krezel, A.M., McNeill, H., Solnica-Krezel, L., Atypical Cadherin Dachsous1b Interacts with Ttc28 and Aurora B to Control Microtubule Dynamics in Embryonic Cleavages, 376-391.e5, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell