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. 2021 Dec 23;10:e74336. doi: 10.7554/eLife.74336

Figure 7. POU-IV-binding motifs are conserved across Cnidaria and Bilateria.

Figure 7.

(A) Motifs enriched in Nematostella vectensis POU-IV chromatin immunoprecipitation sequencing (ChIP-seq) peaks. (B) Homo sapiens POU motifs resulting from sequence alignment and comparison against the Jaspar database. The p-value reported corresponds to the highest p-value for any of the three N. vectensis POU4 motifs found. (C) Electrophoretic mobility shift assay (EMSA) using purified N. vectensis POU-IV protein and a 50 bp DNA probe containing the conserved core motif CATTATTAAT. Note that retardation of probe migration occurs in the presence of POU-IV protein (blue arrowhead; lane 2), indicative of formation of the protein-DNA complex. Retardation is inhibited in the presence of an unlabeled competitor probe (‘specific competitor’; lane 3). Removal of the motif sequence in the probe (‘probe without motif’) abolishes retardation of probe migration by POU-IV (lane 4), demonstrating that the motif is necessary for formation of the protein-DNA complex. The mobility of the probe is further decreased in the presence of the anti-POU-IV antibody (purple arrowhead; lane 5), confirming that the protein bound to the probe is POU-IV.

Figure 7—source data 1. List of 12,972 genome-wide-binding sites for POU-IV.
Figure 7—source data 2. An original gel image used to generate Figure 7C and the original image with relevant lanes labeled.