Figure 4

Smed-2C4 labels multiple cells with distinct morphologies and anatomical locations. (A-F) Whole-mount staining of intact planarians or regenerating planarians with 2C4 (green) and with either 6G10 (magenta) in panels E and F or Lens culinaris agglutinin lectin (LCA, magenta) in panels G and H. (A) 2C4 labels multiple distinct cell types in the intact worm. Closed arrowhead indicates an example of a 2C4-S cell. Open arrowhead highlights a 2C4-N cell. Arrow indicates a large round 2C4-I cell. (B, C) 2C4 labels the anterior and posterior blastema during regeneration in 2, 3, 4, 5, and 7 dpa trunk regenerates. Higher magnification images of the anterior blastemas are shown in C. Arrowhead highlights an example of the 2C4-S cells seen throughout regeneration. (D) Magnified image of 2C4-N cells. Arrows denote the large cell bodies and arrowheads indicate their projections. (E, F) 2C4-I cells are located within the anterior (shown in E) and posterior (shown in F) intestinal branches (delineated by labeling of the intestinal wall musculature with 6G10). (G, H) 2C4 is expressed in a subset of goblet cells marked with LCA. Strongly labeled LCA⁺ cells immediately anterior to the pharynx were weakly labeled with 2C4 (shown in G). In contrast, strongly labeled 2C4 cells in anterior secondary branches were weakly labeled with LCA (shown in H). Dashed boxes in E-G indicate the areas shown in the inset images. Images are maximum intensity projections of optical sections except for A and B. Anterior is to the left in A and to the top in B-G. Image D was acquired adjacent to the pharynx. Scale bars: (A, B) 200 μm; (C, E-H) 50 μm; (D) 20 μm.