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. 2013 Oct 3;9(10):e1003811. doi: 10.1371/journal.pgen.1003811

Figure 3. Functional dissection of CNE1 enhancer activity.

Figure 3

(A) Schematic outlining the organisation of motifs within CNE1. (B, B′) CNE1 transient transgenic zebrafish embryos recapitulate pax3a expression across the AP axis of the CNS, including the spinal cord (n = 66/67). (C) Zebrafish CNE1 sequence specifically drives transgene expression in the Pax3/7 domain of the chick spinal cord, despite widespread transfection of LacZ across the DV axis of the tissue (n = 10/12). (D) Matrix representing the degree of Motif1 conservation across 12 vertebrate genomes. (E, E′) Deletion of Motif1 results in a complete loss of CNE1 activity in zebrafish spinal cord at trunk level (n = 0/22, p<0.0001), however the enhancer remains active in the anterior CNS (n = 22/22) and the most posterior region of the spinal cord (n = 8/22). (F) Loss of Motif1 greatly reduces CNE1 activity in the chick neural tube (n = 1/8, p = 0.0019). Motif2 (G), is not required for CNE1 activity in the zebrafish (H, H′) (n = 37/38) or chick (I) (n = 4/6) spinal cord. Loss of Motif 3 (J) reduces CNE1 mediated transcription across the AP axis of the zebrafish CNS (K, K′) (n = 0/26, p<0.0001) and precludes activity in the chick spinal cord (L) (n = 0/5, p = 0.003). Deletion of Motif4 (M) does not significantly alter the activity of CNE1 in zebrafish (N, N′) (n = 20) or chick (O) (n = 4/5).