Fig. 6. Differential effects of Lhx2 and Lhx5 on the OV and genetic interactions between Lhx1 and Lhx5.(A) Phylogenetic tree of chicken LIM-homeodomain.

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Lhx2, a LIM homeobox gene, is required for eye, forebrain, and definitive erythrocyte development Introduction Lhx2 is a member of the LIM homeodomain.

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Fig. 6. Differential effects of Lhx2 and Lhx5 on the OV and genetic interactions between Lhx1 and Lhx5.(A) Phylogenetic tree of chicken LIM-homeodomain (LIM-HD) factors. Differential effects of Lhx2 and Lhx5 on the OV and genetic interactions between Lhx1 and Lhx5.(A) Phylogenetic tree of chicken LIM-homeodomain (LIM-HD) factors. The LIM-HD factors are depicted by the NJ (neighbor-joining) method. Chicken LMO3 was used as an outgroup. The scale bar is calibrated in substitutions per site. Numbers show bootstrap confidence values. Amino acid sequences used in the tree construction were deduced from the nucleotide sequences listed in supplementary material Table S2. (B–D) Lhx2 is expressed in the forebrain and whole OV at stage 11. Dorsal (B) and lateral (C) views. (D) Transverse section of the OV. (E–H) EGFP fluorescence images at 24 hours post-electroporation. Overexpression of Lhx2 (F) did not induce transformation of the outer optic cup to the NR, whereas overexpression of Lhx5 (G) or Lhx1 (H) induced a protrusion of the OV. (I,J) Histology of an Lhx5-overexpressing retina at stage 24. A second NR transformed from the outer layer of the optic cup. (K,L) In situ hybridization of an Lhx5-overexpressing retina at stage 21. Arrows show activation of Chx10 (K) and suppression of Otx2 (L) in the developing second NR. (M–Q) Whole mount in situ hybridization (WISH) of embryos at Hamburger's and Hamilton's stages 9 (M,N), stage 10 (O,P), and stage 11 (Q). (M,O,Q) Dorsal views. (N,P) Lateral views. Transverse sections cut after WISH are shown in (Q-a,Q-m,Q-p). At the middle OV level (Q-m), Lhx5 is distinctly expressed in the proximal region of the OV, while it is expressed in the ventral diencephalon at the posterior OV level (Q-p). (R–T) EGFP fluorescence (R) and in situ hybridization of Lhx1 (S,T) at 24 hours post-electroporation. Lhx1 expression is ectopically induced in the protruding outer layer of the optic cup after Lhx5 overexpression (arrowhead in S,T). Scale bars: 100 µm. Takumi Kawaue et al. Biology Open 2012;1:1083-1093 © 2012. Published by The Company of Biologists Ltd