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Fig. 3 Lgl2 is required for normal left-right asymmetry. (A) Asymmetric heart looping at 2 dpf visualized in Tg(cmlc2:GFP) transgenic embryos that express GFP in the heart. In control embryos, the heart (black arrow) looped to the right, whereas Lgl2 MO knockdown often resulted in reversed or no heart looping (red arrows). (B) Percentage of embryos with heart-looping defects. Injecting a lower dose of Lgl2 MO-1 revealed that looping defects are dose dependent, and co-injecting lgl2 mRNA partially rescued Lgl2 MO-1 and Lgl2 MO-2 heart defects. Injecting lgl2 mRNA alone also disrupted heart looping. (C) RNA in situ hybridization analysis of spaw expression, which was detected in left lateral plate mesoderm (LPM) (arrowhead) and bilaterally in the tail (arrows) at 14 hpf in controls. Lgl2 MO embryos showed disrupted LPM spaw expression, including right-sided, bilateral and absent expression (red arrowheads). (D) Analysis of LPM spaw expression in control and Lgl2 MO embryos. n, number of embryos analyzed. L, left; R, right.