Figure S3. Validation of Otx1 morpholino knock-down and Otx1 ChIP. Related to Figures 2 and 3. (A) Sequence of the X. tropicalis otx1 ATG region, along with the translation blocking morpholino and the HA-otx1 rescue construct. Underlined on the WT otx1 sequence is the translation start site. In red in the HA-otx1 sequence are 14/25 mismatching bases between WT and rescue sequence that is targeted by the morpholino. (B) RT-qPCR at mid-gastrula of morpholino injected compared to uninjected vegetal masses to test morpholino specificity. dicer1 was used for normalization. Error bars represent variation from three technical replicates. (C) X. tropicalis Otx1 protein sequence where the underlined sequence is the target of the Otx1 polyclonal peptide antibody. In colors are conserved regions of the protein: the homeodomain (orange), the WSP domain (green) and the Otx-tail (red). (D) ChIP-qPCR using the antibody assaying for Otx1 binding in the promoter regions of mesendodermal versus control genes. Error bars represent variation from three technical replicates.
Image published in: Paraiso KD et al. (2019)
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