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.
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.
In the head of both males and hermaphrodites, the expression of the Ptbb-4::YFP transcriptional reporter was observed in many amphid neurons, likely ADL, AFD, ASE, ASG, ASH, ASJ, ASK, ASI, AWA, AWB, and AWC as judged by co-labeling with lipophilic dyes and/or cellular morphology. Other head neurons expressing Ptbb-4::YFP include the cephalic CEP neurons, the outer labial quadrant OLQ neurons, and the mechanosensory FLP neurons. In the midbody and tail, Ptbb-4::YFP expression was found in the PDE postdeirid neurons, PQR (a sensory neuron whose cilium is exposed to the pseudocoelomic fluid), and the phasmid neurons PHA and PHB. Ptbb-4::YFP was also expressed in all 18 ray neurons in the male, consistent with a previous report (Portman and Emmons 2004). Because of abundant ray neuron expression, Ptbb-4::YFP expression in male phasmid and hook neurons could not be reliably confirmed, although expression in these cells is not unlikely. We also observed Ptbb-4::YFP expression in the hermaphrodite HSN motor neurons, the vulval muscles, and several unidentified nonsensory neurons. Expression was also occasionally apparent in the amphid sheath (AMsh) and socket (AMso) glial cells.
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
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