daf-11 : egl-4

The AWC neurons enable C. elegans to sense various volatile attractive odorants. After prolonged exposure to one of these odors, the protein kinase G EGL-4 will translocate from the cytoplasm to the nucleus of the AWC. However, GFP::EGL-4 is constitutively nuclear in guanylyl cyclase mutant daf-11 animals.

dat-1 : paraquat

"We examined whether tunicamycin-induced ER stress will affect these neurons in our assay. The analysis of dat-1::YFP reporter revealed visible defects in neuronal morphology in treated animals illustrated by dendritic/axonal abnormalities and loss of cell bodies (Fig. S1A). A similar result was obtained following PQ treatment (Fig. S1B)"

Tunicamycin : dat-1

"We examined whether tunicamycin-induced ER stress will affect these neurons in our assay. The analysis of dat-1::YFP reporter revealed visible defects in neuronal morphology in treated animals illustrated by dendritic/axonal abnormalities and loss of cell bodies (Fig. S1A). A similar result was obtained following PQ treatment (Fig. S1B)"

daf-2 : odr-10

Reduction of odr-10 levels by RNAi increased daf-2 motility on bacteria significantly, suppressing the decreased velocity seen on bacteria in the daf-2(e1370) mutant alone. daf-2 mutants will stop moving after 1.5 to 3 min when placed onto bacteria, but with reduction of odr-10 levels by RNAi, worms continue to move constantly over a 10 min assay period.

egl-20 : lin-17

"To test whether LRP-2 acts downstream of EGL-20, we ectopically expressed egl-20 from the anchor cell in a lin-17(n671); lrp-2(gk272) double mutant background and compared it to a lin-17(n671); lrp-2(+) strain. If LRP-2 acts downstream of EGL-20, then anteriorly-expressed EGL-20 will not be able to suppress the lin-17 phenotype, with is the result observed (Table 1)."

let-99 : par-3

In one-cell par-3 embryos, LET-99 was present around the entire periphery in all prophase through metaphase stage embryos. The staining intensity at the periphery was comparable with that of the posterior band of wild-type embryos. During anaphase, the LET-99 signal at the anterior and posterior of one-cell par-3 embryos diminished, leaving a band of LET-99. However, in contrast to wild type, this band was symmetrically positioned and will be referredto as the central band.

mec-3 : unc-86

The sister of the PVDR cell will express mec-3-lacZ response to jeEx100 (unc-86+++). At a low frequency (3%), an extra mec-3-expressing cell adjacent to PVDR is seen in a ced(+) ; jeEx100 strain. When this occurs, there are five neuronal nuclei in the PVDR cluster, suggesting a transformation into a mec-3-expressing cell of the PVDR sister, which normally undergoes programmed cell death. When cell death is blocked with a ced-4(n1162) mutation, the frequency of ectopic mec-3-LacZ cells increases to 41%, indicating that in the presence of a high unc-86 gene dose, the PVDR sister can often express mec-3-LacZ if it does not first undergo programmed cell death .

tph-1 : unc-2

Temperature shift experiments were performed using unc-2;daf-4(m592) double mutants to identify the developmental period during which thermal enhancement of tph-1::GFP expression requires UNC-2 regulation of TGF-b signaling. Animals grown to the L3 larval stage or beyond at 15 Centigrades did not develop enhanced expression of the tph-1::GFP transgene when shifted to 25 Centigrades and, conversely, animals grown to the L3 larval stage at 25 C retained high expression of the tph-1::GFP transgene in the ADF neurons when downshifted to 15 Centigrades. This experiment demonstrates a developmental window during the L2 larval stage, around the time at which the decision is made to commit to dauer larva development, beyond which thermal inactivation of DAF-4 will not change the percentage of ADF neurons with increased expression of the tph-1::GFP transgene. Together these data show that UNC-2 and the TGF-b pathway transduce a developmental thermal cue regulating expression of tph-1.

lin-39 : nhr-43

When RNAi was performed on worms carrying the lin-39::GFP transcriptional reporter to reduce nhr-43 function, their progeny showed a decrease in the number of animals expressing wild type levels of GFP in the vulval precursor cells at the L3 stage. qRT-PCR on nhr-43(tm1381) mutant animals at the L3 stage also showed a decrease in lin-39 expression compared to control animals. Authors also found that fewer embryos derived from hermaphrodites treated with nhr-43 RNAi showed lin-39::GFP expression in P5 - 8 compared to control animals, indicating that nhr-43 positively regulates lin-39::GFP in the embryonic P cells. This embryonic defect could explain the reduction in GFP expression in the VPCs in the larva, however a decrease in lin- 39::GFP expression was also observed if nhr-43 RNAi was performed on L1 worms and then the same animals were observed in the L3 stage (51% of nhr-43(L1 RNAi) animals showed wild type expression in all VPCs versus 76% of RNAi control, p < 0.001). In summary, orphan nuclear hormone receptor NHR-43 binds to a site located far upstream of the lin-39 initiation codon, and nhr-43 function is required for wild type levels of lin-39 expression in the embryo and larva in cells that will participate in vulval development.