Fig. 3. Genetic interactions between unc-53 and unc-6 and between unc-53 and unc-5 affect intracellular UNC-40::GFP localization.(A–H) Photomicrographs.

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Fig. 3. Genetic interactions between unc-53 and unc-6 and between unc-53 and unc-5 affect intracellular UNC-40::GFP localization.(A–H) Photomicrographs of the localization of UNC-40::GFP in the HSN neuron of L2 stage larvae. Genetic interactions between unc-53 and unc-6 and between unc-53 and unc-5 affect intracellular UNC-40::GFP localization.(A–H) Photomicrographs of the localization of UNC-40::GFP in the HSN neuron of L2 stage larvae. Ventral is down and anterior is to the left. Scale bar: 5 µm. UNC-40::GFP is ventrally localized in the wild-type animals (A), but in unc-6(ev400) mutants it is evenly distributed (B). Although UNC-40::GFP is ventralized in unc-53(n152) mutants (C), in unc-53(n152); unc-6(ev400) mutants it is polarized to different sides, including dorsally (D). Although UNC-40::GFP is ventralized in unc-5(e53) mutants (E), in unc-53(n152); unc-5(e53)mutants it is more evenly distributed (F) and is similar to localization in unc-6(ev400) mutants. (G) Graph indicating the dorsal–ventral orientation of UNC-40::GFP. The graph shows the average ratio of dorsal-to-ventral intensity from linescan intensity plots of the UNC-40::GFP signal around the periphery of the HSN cell. Wild-type animals show a strong ventral bias, whereas there is a uniform distribution in unc-6(−) mutants and an increased dorsal bias in unc-53(n152); unc-6(ev400) mutants. (*) statistic difference (P<0.05, one-tailed Student's t-test); (n.s.), statistically, no significance. (H) Graph indicating the anterior–posterior orientation of UNC-40::GFP. To determine orientation, line-scan intensity plots of the UNC-40::GFP signal across the dorsal periphery of the HSN cell were made, the dorsal surface was geometrically divided into three equal segments, and the total intensity of each was recorded. The effect of the mutations on UNC-40 distribution can be accessed by comparing the two graphs, for example in unc-6(ev400) mutants UNC-40 is evenly distributed as indicated by the equal dorsal-to-ventral ratio and the uniform anterior–posterior value. In contrast, UNC-40 is polarized to different sides in unc-53(n152); unc-5(e53)mutants as indicated by the increased dorsal-to-ventral ratio and the increased bias to localize at the middle of the dorsal side (red) or to the anterior (blue) or posterior (green) sides. Gauri Kulkarni et al. Biology Open 2013;2:1300-1312 © 2013. Published by The Company of Biologists Ltd