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. 2011 Oct 13;7(10):e1002308. doi: 10.1371/journal.pgen.1002308

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Yamamoto et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A–F) The expression of cwn-1p::CWN-1::Venus (A), cwn-2p::CWN-2::Venus (B, C), ceh-22p::CWN-1::Venus (D), and ceh-22p::CWN-2::Venus (E). Anterior is to the left and ventral to the bottom, except for (B), in which anterior is to the upper-left and ventral toward the right. Scale bars: 20 µm. Confocal images in (A, B) and (D, E), were focused on hypodermal cells. The focus is on the pharynx/intestine in (C). (A, B) Dashed lines outline seam cell nuclei. White arrowheads and blue arrows indicate some of the GFP puncta and granule autofluorescence, respectively, which were observed even in the corresponding DIC images. Insets show the area indicated by orange dashed boxes, magnified 2-fold; they illustrate the efficient diffusion of cwn-2p::CWN-2::Venus but not of cwn-1p::CWN-1::Venus. (C) Expression of cwn-2p::CWN-2::Venus in the pharynx and near the intestine. Orange, white, and blue lines outline the pharynx, intestine, and gonad, respectively. Puncta of cwn-2p::CWN-2::Venus near the intestine (white arrowheads) were observed only in the anterior region. Puncta in the intestine are gut granule autofluorescence. (D, E) Diffusion of ceh-22p::CWN-2::Venus (E) but not ceh-22p::CWN-1::Venus (D) was observed in the anterior hypodermis near the pharynx. White arrow heads, blue arrows and the orange arrow indicate GFP puncta, hypodermal cells expressing the elt-3::GFP marker, and neuronal processes expressing the mec-4::GFP marker, respectively. (F–H) Each colored box represents the polarity of individual seam cell divisions, as in Figure 1C. (F) The egl-20 phenotype was not affected by EGL-20 expression in the M cell (hlh-8p::EGL-20). Since the M cell is on the right side of the animals, we scored seam cells only on the right side. (G) cwn-1p::CWN-1::Venus rescued the defect of the cwn-1 mutation in cwn-1; egl-20. (H) The phenotype of cwn-1; egl-20 cwn-2 was rescued by cwn-2p::CWN-2::Venus, by CWN-2 expressed in the pharynx (ceh-22p::CWN-2::Venus) or in the cells near the anus (egl-20p::CWN-2::Venus#1 and #2 with mec-4::GFP and egl-5::GFP as coinjection markers, respectively), or by CWN-1 expressed in the pharynx (ceh-22p::CWN-1::Venus).