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.
CLC-2-GFP was detected in hypodermal seam cells, which fuse to form a single elongated syncytium. However, in contrast to CLC-1-GFP, under the expression condition used in this study, GFP signals did not appear to be concentrated at the borders between the seam cell syncytium and surrounding hypodermal cells. When adult wild-type worms were whole-mount stained with anti-CLC-2 pAb, the seam cell syncytium appeared to be outlined to give two parallel lines, although these lines were thick and not very sharp.
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
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