Figure 2. Knock-down of hccs in medaka recapitulates the phenotypic features of MLS syndrome.

• A–E. Bright-field dorsal (A–C) and lateral (D, E) views of wt (A), hccs-MO (B, E) and control-MO (C, D) -injected embryos at st38. hccs-morphants display microphthalmia (vertical dashed line in B), microcephaly (horizontal dashed line in B) and cardiac defects including failure of heart loop formation and pericardial oedema (red arrow in E). In a significant number of the microphthalmic embryos (50%), RPE layering and closure of ventral optic fissure were also impaired resulting in coloboma formation (black arrow in E; see also Supporting Information Table S1). Embryos injected with control-MO did not show any abnormal phenotype (C, D). a, atrium; v, ventricle. Scale bars: 100 µm.
• F. Analysis of the eye size at st24 and st38 (error bars are SEM; n ≥ 19 eyes, p-values were calculated by two-tailed Student's t-test).
• G. Western blotting analysis revealed decreased levels of total Cytc in morphants (hccs-MO) at st19 and st24 compared to control embryos (control-MO). GAPDH was used as loading control.