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. 2022 May 27;50(10):5652–5671. doi: 10.1093/nar/gkac408

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — The TOP-3-ZnF domain influences localization of the BTR complex. (A) Super-resolution images of gfp::rmh-1; top-3::ollas (left) and him-6::ha; top-3::ollas (right) nuclei in mid pachynema (top) and late pachynema (bottom). Nuclei were stained with anti-SYP-1 (blue), anti-GFP (yellow) and anti-OLLAS (magenta) antibodies (left) and with anti-SYP-1 (blue), anti-HA (cyan) and anti-OLLAS (magenta) antibodies (right). Half part of the nucleus was projected. Insets (top, right) show magnified boxed regions, highlighting co-localization of the proteins. (B) TOP-3::OLLAS localization in mid pachynema (top) and late pachynema (bottom) in the indicated genotypes. Gonads were stained with DAPI (cyan) and anti-OLLAS antibody (red). (C) HIM-6::HA localization in mid pachynema (top) and late pachynema (bottom) in the indicated genotypes. Gonads were stained with DAPI (blue) and anti-HA antibody (yellow). (D) Upper panel: GFP::RMH-1 localization in mid pachynema (top) and late pachynema (bottom) in the indicated genotypes. Gonads were stained with DAPI (magenta) and anti-GFP antibody (green). Lower panel: quantification of GFP::RMH-1 foci in the meiotic region of the C. elegans gonad. The scatter plot indicates the number of foci per nucleus: zone 1 = transition zone; zone 2 = early pachynema; zone 3 = mid pachynema; zone 4 = late pachynema. For each genotype, the mean ± SD number of foci in each zone are indicated, with n = the number of nuclei assessed. gfp::rmh-1: zone 1, 1.2 ± 1.2, n = 111; zone 2, 15.22 ± 6.9, n = 155; zone 3, 18.64 ± 6.1, n = 44; and zone 4, 8.3 ± 2.1, n = 92. gfp::rmh-1; top-3-ZnF: zone 1, 1.1 ± 1.3, n = 103; zone 2, 3.6 ± 3.8, n = 167; zone 3, 11.4 ± 4.7, n = 186; and zone 4, 7.7 ± 1.7, n = 114. gfp::rmh-1; top-3-ZnF; rmh-2: zone 1, 0.4 ± 0.7, n = 83; zone 2, 1.6 ± 3, n = 89; zone 3, 6.3 ± 3, n = 62; and zone 4, 6.6 ± 1.6, n = 47. **** P < 0.0001, as determined using the Mann–Whitney test; non-significant differences are not shown. (See Supplementary table 1 for more details of the statistical analysis).