Organ left-right lateral defect in embryos treated with different ways. (A-D, O) The pattern of heart displayed in controls and aplnra/b morphants. Compared with controls (A, 98.6%, n=72), embryos injected with aplnra MO displayed normal-loop (B, 62.5%, n=112; p<0.01), reversed-loop (C, 17.8%, n=112; p<0.01) and no loop (D, 19.6%, n=112; p<0.01). (O, column 1 to 4) The cartogram of heart LR defect for embryos treated with different MOs or mRNA. Only more than 18.2% of aplnrb morphants displayed heart LR defect (O, column 4, n=115; p<0.05). Aplnra mRNA injection (O, column 3, 26.8%, n=71; p<0.05) partially rescued the heart LR defect in aplnra morphants (O, column 2, 37.5%, n=112). (E-J, P). While 97.3% of control embryos showed left-sided expression of cp (E, n=112), 18.2.0% (F, n=176, p<0.01), 9.1% (G, n=176, p<0.05) and 10.8% (P, n=176, p<0.05) of aplnra morphants showed both sided and right-sided expression of cp, or disappear. (H-J, P). Compared with that in aplnra morphants, aplnra mRNA partially rescued liver LR defect (24%, n=121, p<0.05). In aplnra/b double morphants, the expression of cp was greatly downregulated (100%, n=195, p<0.001), and more embryos displayed liver LR defect (H-J, P, 51.3%, n=195, p<0.05) than that in aplnra morphants (P, 38.1%, n=176). In embryos injected with aplnra sgRNAs and Cas9 protein, 15.8%; and 6.4% of them displayed right sided and both sided/middle liver (P, n=233, p<0.05). (K-N, Q). otx5 was expressed in the middle telencephalon. All the embryos injected with Cont MO displayed the expression of otx5 in the middle telencephalon (K, 100%, n=32, and Q, column 1).While 61.7% of embryos injected with aplnrb MO showed both-sided otx5 in telencephalon (M, Q, n=34), only 5.5% of aplnra morphants displayed both-sided otx5 (Q, 37). For observation, heart, ventral view; cp, spaw and otx5, dorsal view.
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