Effect of <i>sun-1</i> phosphosite mutations on the duration of meiotic stages, DSB turnover, SUN-1 aggregates, chromosome movement, and synapsis.

<p>(A) TZ nuclei of <i>sun-1(wt); sun-1(ok1282</i>) (top), and <i>sun-1(allA); sun-1(ok1282)</i> (bottom) hermaphrodite gonads stained with DAPI (left; blue in merged picture) and anti-GFP (middle; green in merged picture). Scale bars, 10 µm. (B) First frame of <i>in vivo</i> time-lapse GFP-recorded TZ nuclei of <i>sun-1(wt); sun-1(ok1282)</i> (top, left) and <i>sun-1(allA); sun-1(ok1282)</i> (bottom, left) hermaphrodite gonads. Displacement tracks of SUN-1 aggregates represent 2D plotted chromosome end movements over 3 min (right). Average speed of SUN-1 aggregates in TZ (<i>n</i> = 9 nuclei, followed over 3 mins). Scale bar, 2 µm. (C) Schematic of a wild-type hermaphrodite gonad with nuclei in the corresponding zones, as used in the quantifications in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003335#pgen-1003335-g003" target="_blank">Figure 3D</a>: chromatin (blue) and SUN-1 morphology (green). Gonad is subdivided into ten zones, as used for the RAD-51 foci quantification in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003335#pgen-1003335-g003" target="_blank">Figure 3E</a>. Meiotic entry and beginning of cellularization are indicated. (D) Representation of relative time of residency in different meiotic stages, as assessed by presence of SUN-1 aggregates in different mutant backgrounds, normalized to the length of the gonad (from meiotic entry [0%] to start of meiocyte cellularization at diplotene [100%]). Nuclei were sorted into three categories: “TZ” (dark green, more than one SUN-1 aggregate), “early pachytene” (light green, one SUN-1 aggregate), and “late pachytene” (red, no SUN-1 aggregate). Categories were assigned once ≥50% of nuclei in a cell row fulfilled one of these criteria. At least eight gonads were counted per genotype. (E) Average numbers of RAD-51 foci per nucleus in the hermaphrodite gonad of different <i>sun-1</i> phosphosite mutants. Gonads were divided into ten zones, as schematically indicated in (B). Three gonads per genotype were counted. (F) Nuclei from early and late pachytene zones of <i>sun-1(wt); sun-1(ok1282)</i> (top) and <i>sun-1(allA); sun-1(ok1282)</i> (bottom) hermaphrodite gonads stained with anti-HTP-3 (left, green in merged) and anti-SPY-1 (middle, red in merged). Scale bars, 10 µm.