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. 2022 Nov 10;13:6805. doi: 10.1038/s41467-022-34450-y

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Overall structure of the full-length LPD-3 assembled from four segments whose structures were separately predicted by AlphaFold v2.0, with both N and C-termini noted. b Cross-sectional view of the LPD-3 structure showing the hydrophobic tunnel running continuously along the entire inside. c Ribbon representation of the N-terminal part of LPD-3, with the G200E mutation indicated in yellow. d Structure of the LPD-3 C-terminus showing an amphiphilic patch (hydrophobic, red; hydrophilic, blue) that was used to generate mCherry-fused LPD-3 reporters. e Representative fluorescence images showing co-localization (arrow heads) of ER membrane markers with a mCherry fusion reporter of the LPD-3 N-terminus (rpl-28p::LPD-3Nt::mCherry with LPD-3 a.a. 1-72, unc-54p::GFP as co-injection marker). f Representative fluorescence images (from two independent transgenic extrachromosomal arrays #1 and #2) showing co-localization (arrow heads) of Akt-PH::GFP that binds to PM-PIP3 with a mCherry fusion reporter (ges-1p::mCherry::LPD-3Ct, a.a. 3945-4018) of the LPD-3 C-terminus. g Representative confocal fluorescence images showing endogenous LPD-3::GFP (generated by CRISPR/Cas9-mediated knock-in and split GFP complementation) in apposition with mScarlet::SP-12(ER). h Representative confocal fluorescence images showing endogenous LPD-3::GFP that co-localizes (arrow heads) with mScarlet::MAPPER, a marker for ER-PM junctions. Scale bar, 50 µm.