Endogenous DVC1 was predominantly expressed during S and G2 phases of the cell cycle. Upon exit from mitosis, DVC1 expression was rapidly downregulated with kinetics similar to that of known APC-Cdh1 targets such as cyclin A, and reaccumulated as cells entered S phase.
Similar to UNC-11 antibody staining, UNC-11::GFP expression appeared pan-neuronal and was also observed within coelomocytes. Not described by Nonet el al 1999, we observed GFP puncta in apical epithelial tissue just below the cuticle surface in both the head and in the body, an accumulation of UNC-11::GFP at the plasma membrane in the seam cells and weak staining in the vulval precursor cells (VPCs) at the interface between the VPCs and the vulval lumen. Finally, Nonet et al. described diffuse staining within the intestine whereas we did not observe detectable levels of intestinal expression, punctate or otherwise.
To determine in which cell(s) tiar-2 functions, we constructed a GFP-TIAR-2 mini-gene translational reporter and observed broad expression in many cell types, including mechanosensory neurons.
HSP-4::GFP was exclusively localized in the PVD soma, colocalizing with a rough ER marker TRAM, HSP-4's endoplasmic reticulum localization pattern is consistent with the observation that its mammalian homolog BiP is localized in rough endoplasmic reticulum.
Time-lapse fluorescence microscopy was performed, including DIC for morphology. Gene expression patterns were summarized in 4 manners: Average over time, Average over time and at different positions along the anterior-posterior (AP) axis, a voxelized representation over time, and on individual cells overlaid from a reference coordinate dataset (https://doi.org/10.1016/j.ydbio.2009.06.014). The analysis was done with a pipeline based on the multi-purpose image analysis software Endrov (https://doi.org/10.1038/nmeth.2478), which further is needed to browse the raw recording data. Thumbnail movies were also generated, using maximum Z projection for the 3D fluorescence channel. Raw recordings available in the Endrov OST-file format are available at https://www.ebi.ac.uk/biostudies/studies/S-BIAD191?query=S-BIAD191