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. 2018 Oct 31;14(10):e1007776. doi: 10.1371/journal.pgen.1007776

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© 2018 Nguyen et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 9 — (A) Representative images of wild-type and zhp-4(vv96) diakinesis nuclei stained with α-AIR-2 (green) and α-HTP-3 (red). Wild-type bivalents exhibit the axial element HTP-3 in a cruciform shape (defining both the long and short axial arms) while AIR-2 is restricted to the short arm. AIR-2 localization in zhp-4(vv96) mutants is variable and it can be detected on one HTP-3 axis of a well-formed bivalent structure (bivalent), or be undetectable on an aberrant structure (tethered). Scale bars 1 μm. (B) SC component SYP-1 (red) and HTP-3 (green) immunolocalization in diakinesis nuclei showing representative bivalent structures. SYP-1 localization is restricted to the short arms of the bivalent in wild-type and zhp-4(H26A) mutants, but is detected along both long and short arms in the zhp-4(vv96) mutants. Scale bars 1 μm. (C) α-HTP-1 (green) and α-HTP-3 (red) staining of diakinesis nuclei showing axis component HTP-1 restriction to the long arm of the wild-type and zhp-4 ring mutant bivalent while it is continued association with the short and long arm of the bivalents in zhp-4(vv96) mutants. Scale bars 1 μm. (D) -1 diakinesis oocyte (the last oocyte of the germline prior to entry into the spermatheca) stained with α-SMO-1 (green; the C. elegans SUMO ortholog) and α-HTP-3 (red), with magnified insets of individual chromosomal figures from individuals of the indicated genotypes (1–5). SMO-1 localizes to the short arms of bivalents in wild types, a pattern dependent on CO formation since SMO-1 remained associated with chromatin in spo-11(ok79) mutants. In both zhp-4(vv103) and zhp-4(vv96) nuclei, SMO-1 remained associated with the chromatin, even on bivalents assessed by the volume of the chromosome structures and HTP-3 staining. In zhp-4(H26A) mutants, SMO-1 localizes to the short arms axis of all bivalents and remains with the chromatin of the univalents. Magnifications of in lower row show (1) wild-type bivalent with SMO-1 localized to the short arms, (2) a bivalent in zhp-4(vv103) mutants with SMO-1 localized to the chromatin, (3) well-formed bivalent (as assessed by HTP-3 staining) in zhp-4(vv96) mutants without SMO-1 on the short arms, (4) example of tethered bivalent with enriched localization of HTP-3 at the tether and persistent chromatin localization of SMO-1, (5) a univalent in spo-11(ok79) mutants exhibiting SMO-1 localization with chromatin. Scale bars for main panel 5 μm; bivalent magnifications (1–5) 1 μm.