Fig. 5.

The expression of the Netrin receptor UNC-40 is downregulated in daf-16 mutant. (A) The DAF-16 binding sequence in the promoter (yellow box) of the unc-40 gene is highlighted. (B,C) The percentage of ‘ventral targeting’ events (B) and the RI values (C) at 24 h post-axotomy in the unc-40(lf) and unc-6(lf) backgrounds with or without the daf-16f transgenes expressed under touch neuron-specific promoter Pmec-4 [shrEx436] or muscle-specific promoter Pmyo-3 [shrEx437]. Similar data were also shown in akt-1(lf) background with or without the mutations in unc-6 or unc-40 genes. N (independent replicates)=2-6. (D,E) The confocal images of PLM cell body (D) and PLM axon (E) co-expressing the Punc-40::UNC-40::GFP [icIs132] and Pmec-4::mScarlet [shrEx209] in wild type (WT) and daf-16(lf) before and after axotomy. For cell body, a single z-plane is shown (D). Two to four z-planes were projected to show the tip of the regrowing axon (E). The observations for E are illustrated on the right side. The yellow dotted ROIs on the cell body and axon were used to measure the intensity of UNC-40::GFP and mScarlet before and after axotomy. The red arrows represent the site of injury. The black arrows and arrowheads represent the PLM axon and other UNC-40::GFP-expressing cells, respectively. (F) The mean intensity of UNC-40::GFP normalized to mScarlet from the ROI on the cell body. N=3-8. (G) The mean intensities of UNC-40::GFP normalized to mScarlet from the ROI on axon. N=3-8. Data are mean±s.d. *P<0.05; **P<0.01; ***P<0.001; ns, non-significant [Fisher's exact test (B); ANOVA with Tukey's multiple comparisons test (C,F,G)].