Fig. 9

Synergistic effect of sox4 and sox11 knockdown on coloboma. (A–H) Representative images of embryos injected with control MO (A, E), a half-dose of sox4 MO (B, F), a half-dose of sox11 MO (C, G), and half-doses of sox4 and sox11 MOs (D, H). (A, E) Half the normal dose of sox4 MO caused a low incidence of mild coloboma. (C, G) Half the normal dose of sox11 MO resulted in low incidence of coloboma and lens malformations at 24 hpf (C, arrow). (D, H) Injection of a half-dose of both sox4 and sox11 MO significantly increased the incidence of coloboma, and the coloboma phenotype was more severe compared to sox4 or sox11 morphants alone. (I) Quantification of the proportion of embryos with coloboma at 48 hpf (half-dose sox4 MO, 11.05±3.15%, n=16/145; half-dose sox11 MO, 22.31±4.0%, n=16/71; half-dose of both sox4 and sox11 MO, 72.47±2.95%, n=51/70; Fisherós exact test, P=0.0001). (J) Sox11 can compensate for the loss of Sox4. Injection of sox11 mRNA into sox4 morphants significantly reduced the incidence of coloboma from 41.09±7.36% (n=113/275) to 10.14±3.09% (n=28/276; Fisherós exact test, P<0.0001). All scale bars equal 100 µm.