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Fig. 5. nhr-25 RNAi affects development of L4-adult seam epidermis and expression of a cadherin cdh-3::gfp reporter. (A) Direct GFP fluorescence of cdh-3::gfp reveals continuity of the seam and proper formation of the vulva in control L4 larvae. (B,C) Large gaps separate seam cells in worms subjected to nhr-25 RNAi by feeding since hatching (B) or starting at 14 hours after hatching (C). Arrowhead points to a seam cell that failed to elongate in the anteroposterior direction, asterisks mark the positions where the vulva would normally develop. (D,E) Confocal images acquired under identical settings show that cdh-3::gfp expression is reduced or nearly undetectable in nhr-25 dsRNA-fed L4 animals (E) compared with controls (D). (F) Fusion of incorrectly aligned seam cells in late L4 nhr-25(RNAi) larvae (obtained by dsRNA injection of their parents) causes bifurcations and loops in the syncytium, as revealed by cdh-3::gfp. Arrows point to three seam nuclei. (G,H) L4 progeny of adults injected with nhr-25 dsRNA contain superfluous, irregularly spaced seam-cell nuclei marked by scm::gfp (H, arrows) compared with controls (G).

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