We found a similar pulse of NHR-23::mScarlet in L4.3 vulval precursor cells as that reported by Kinney et al..
nhr-23::mScarlet was also detectable in seam and hypodermal cells at this stage, epithelial cells that synthesize cuticular components. NHR-23::mScarlet was detected in pachytene nuclei in L4 animals and its zone of expression became restricted in young adult animals. This expression pattern is again similar to NHR-23::GFP::AID*::3xFLAG (Ragle et al. 2020). However, we also observed some interesting differences compared to NHR-23::GFP::AID*::3xFLAG. In our previous work, we observed that NHR::23::GFP became undetectable in ovulating adults (Ragle et al. 2020). In contrast, NHR-23::mScarlet appears to be diffusely expressed in young adult residual bodies and spermatocytes. This diffuse expression persists into ovulating adults, and appears restricted to the spermatheca. The NHR-23::mScarlet signal is specific to
nhr-23::mScarlet::3xMyc animals as no diffuse expression in the germline is observed in wild-type animals. One can observe the diffuse expression pattern in L4.5 germlines several rows of cells after NHR-23::mScarlet::3xMyc localization is lost from nuclei.