On of percentage of nuclei that migrated commonly, initiated nuclear migration but failed to complete it (partial), or failed to move at all (static). (F) Quantification with the time it took nuclei to reach the dorsal midline in the embryo. Nuclei were categorized into those that reached the midline inside 10 min from the completion of intercalation (green), at one hundred min (orange), at 30 min (blue), or in no way (red). Considerable statistical variations as determined by two contingency tests are noted around the left. (G) The distance a nucleus traveled in the initial ten min immediately after completion of intercalation plotted within a histogram. Every person nucleus was binned into 0.5-m increments.from an extrachromosomal array (Fridolfsson and Starr, 2010) was crossed to unc-84(P91S) and unc-84(null) animals. Embryos in the PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/21269315 stage at which hyp7 nuclear migration would commonly happen had been identified applying DIC microscopy. LMN-1::GFP was then imaged in these embryos at 1-s intervals for 3 min to comply with modifications in nuclear envelope morphology throughout nuclear migration. Films of LMN-1::GFP in CFMTI manufacturer wild-type, unc-84(null), and unc-84(P91S) embryos had been visually distinct (Supplemental Movies S4 6). Nuclei in wildtype embryos underwent big movements–greater than half the width of a nucleus–and have been nearly constantly moving (Figure 5A and Supplemental Movie S4). In contrast, unc-84(null) nuclei tended to remain in location over various minutes of filming (Figure 5B and2858 C. R. Bone et al.Supplemental Movie S5); most movements have been on account of the drift on the entire embryo inside its eggshell. Of interest, in unc-84(P91S) nuclei, each phenotypes were visualized. Some nuclei had been observed undergoing substantial directional movements of up to 1 mmin, whereas other nuclei did not move at all. To categorize the movements of LMN-1::GFP throughout nuclear migration, we created projections combining every frame of an eight min, 20 s time-lapse series (Figure 5, A ). The projections had been split into three colors to show the direction of movement. Magenta signifies the first third from the series, yellow the second, and cyan the final third. Working with the time-lapse projections of LMN-1::GFP, we binned nuclei into three categories according to the size of an individualMolecular Biology from the CellFIGURE five: LMN-1::GFP shows dynamic nuclear morphology for the duration of nuclear migration. (A ) Images of embryos expressing LMN-1::GFP particularly in hypodermal cells in the start of time-lapse imaging. Dorsal views; anterior is left. Insets show the identified nucleus in the beginning (magenta) and end (cyan) on the eight min, 20 s film. Arrows in insets show the path the nucleus is supposed to be moving. (A) Wild-type, (B) unc-84(null), and (C) unc-84(P91S) embryos. (A ) Time projections of 500 frames taken at 1-s intervals. In these projections, frames 166 are colored magenta, 16733 are yellow, and 33400 are cyan to show the direction of movement (A ). A second time-lapse projection from the similar embryo for unc-84(P91S) (C). The arrowheads in C and C mark a unc-84(P91S) nucleus that was migrating normally in time-lapse 1 (C) but then failed to continue migration in time lapse 2 (C). Scale bar, ten m. (D ) Nuclei were classified into three categories: no movement, compact movement, and massive movement. The percentage in each and every category is depicted. Considerable statistical variations as determined by two contingency tests are noted around the left. The arrow inside a is definitely an example of a sizable movement, and also the arrow in B demonstrates no movement.c.
