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. 2020 Oct 22;8:594092. doi: 10.3389/fcell.2020.594092

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Copyright © 2020 González-Arranz, Gardner, Yu, Patel, Heldrich, Santos, Carballo, Jaspersen, Hochwagen and San-Segundo.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of H2A.Z and Ndj1 interaction by bimolecular fluorescence complementation (BiFC) assay. (A,B) H2A.Z and Ndj1 interact at the nuclear periphery. Microscopy fluorescence images of cells expressing HTZ1 fused to the N-terminal half of the Venus^YFP (VN) and/or NDJ1 fused to the C-terminal half of the Venus^YFP (VC) as indicated. Nuclei are stained with DAPI (blue). The reconstitution of Venus^YFP fluorescence resulting from H2A.Z-VN/Ndj1-VC interaction appears in yellow. Images were taken 16 h after meiotic induction. The strains in (A) are: DP1540 (HTZ1-VN), DP1541 (NDJ1-VC), and DP1493 (HTZ1-VN NDJ1-VC swr1Δ). The strains in (B) are: DP1496 (HTZ1-VN NDJ1-VC) and DP1493 (HTZ1-VN NDJ1-VC swr1Δ). (C) H2A.Z and Ndj1 interaction at the nuclear periphery depends on the 2–64 N-terminal domain of Mps3. Mps3-mCherry signal is shown in red, Venus^YFP in green, and DAPI in blue. The cells were imaged 16 h after meiotic induction. The strains in (C) are: DP1511 (MPS3-mCherry HTZ1-VN NDJ1-VC) and DP1512 (mps3-Δ2-64-mCherry HTZ1-VN NDJ1-VC). A single medial plane is shown in (A–C). (D) Quantification of the percentage of cells displaying Venus^YFP fluorescent signal in the experiments shown in (A–C). At least three quantification analyses were performed. The total number of cells scored (n) is shown. Error bars, SD. Scale bar, 2 μm.