Lpar2b regulates the patterning of the pLLP. (A) Confocal images showing the rosette structures in the pLLP of Tg(-8.0cldnb:lynEGFP) embryos at 32 hpf. (B) Immunostaining of ZO1 (red) in the pLLP of Tg(-8.0cldnb:lynEGFP) embryos at 32 hpf. Arrowheads mark the center of rosettes in (A,B). (C) Whole-mount ISH shows the expression of genes as indicated in the pLLP at 30–32 hpf in embryos injected with control or lpar2b MO. (D) Whole-mount ISH detects the expression of pea3 in the pLLP and midbrain-hindbrain boundary (MHB) regions in wild-type (WT) and Tg(hsp70:caFGFR1) embryos at 32 hpf. The Tg(hsp70:caFGFR1) embryos were heat shocked at 24 hpf for 1 h at 37°C. The pLLP is outlined by dots in (C,D). (E) Epifluorescence images showing Tg(-8.0cldnb:lynEGFP) /Tg(hsp70:caFGFR1) double transgenic embryos injected with lpar2b MO. Upper panel, without heat shock; lower panel, with heat shock. Arrowheads, a trail of cells left by pLLP. (F,G) Quantification of the area expressing pea3 (F) or lef1 (G) in the pLLP. **p < 0.01; #p > 0.05 compared to control.