Fig. 4. Immunolocalization of Cs-lam3/4/5. (A-C) Single confocal sections through the notochords of wild-type ascidian embryos stained for Cs-lam3/4/5 at (A) late neurula, (B) early tailbud and (C) late tail stages. Anterior is to the left. Green arrowheads indicate the lateral edges of the notochord or notochord primordium. (D) Tangential section grazing the perinotochordal surface of a wild-type late-tail-stage embryo. (E) Reconstructed z-section across the notochord of a mid-tail wild-type embryo. Red 'm's indicate the lateral blocks of muscle cells. (F) Single confocal section through the notochord of a mid-tail-stage chm/chm embryo. (G-I) Single confocal sections through the notochords of (G) mid-tail, (H) mid-late tail and (I) late tail stage aim/aim embryos. Red arrowheads show ectopic laminin localization on intranotochordal cell surfaces. (J) Reconstructed z-section across the notochord of an aim/aim mid-late-tail-stage embryo. Red 'm's indicate the lateral blocks of muscle cells, and the red arrowhead shows ectopic laminin. (K) Representative images from multiple wild-type embryos stained under standard conditions with the Cs-lam3/4/5 antibody, the Cs-lam3/4/5 antibody blocked by prebinding to Cs-lam3/4/5 peptide, or with control non-immune normal rabbit serum. (L,M) Cs-lam3/4/5 antibody staining (red) and GFP staining (green) in C. intestinalis embryos electroporated with (L) notochord-specific expression plasmids for both Cs-lam3/4/5 and GFP, or (M) GFP plasmid alone.