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Fig. S1
Efficiency and specificity of sox11 morpholinos. (A) Schematic representation of the pEF1α:GFP plasmid containing a portion of the sox11 52 UTR placed upstream of the GFP reporter (top). The binding site for the sox11 morpholino is shown in red. Separate reporters were constructed for the sox11a and sox11b MOs. (Center) Lateral view (anterior at top) of 24 hpf embryos injected with EF1α- sox11a/b-GFP plasmids alone (left) or with both sox11 MOs. No GFP expression was detected in the embryo injected with sox11 MOs. (Bottom) Quantification of the proportion of GFP-positive embryos at 24 hpf. The sox11 MOs were highly effective at blocking GFP expression. Number of embryos analyzed: pEF1α-sox11-GFP plasmid alone, n = 169; pEF1α-sox11-GFP + sox11 MOs, n = 140, 3 independent repeats; *p = 0.004, Student′s t-test. (B) Both sox11a and sox11b contribute to abnormal lens and coloboma phenotypes observed in sox11 morphants. The proportion of embryos displaying either phenotype was significantly higher when injected with sox11a and sox11b MOs simultaneously, compared to either MO alone. Number of embryos analyzed: 24 hpf control MO, n = 463; 2 dpf control MO, n = 441; 24 hpf sox11a MO, n = 229; 2 dpf sox11a MO, n = 214; 24 hpf sox11b MO, n = 341; 2 dpf sox11b MO, n = 316; 24 hpf sox11a + sox11b MO, n = 271; 2 dpf sox11a + sox11b MO, n = 262, 3 independent repeats. *p<0.001, Fisher′s exact test. (C) A second non-overlapping sox11 MO (that targeted both sox11a and sox11b simultaneously) produced the same coloboma phenotype in similar proportion to the first set. Number of embryos analyzed: control MO, n = 186 embryos; sox11 MO, n = 194, 3 independent repeats; *p<0.001, Fisher′s exact test. MO, morpholino; hpf, hours post fertilization; dpf, days post fertilization.