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. 2018 Jun 22;9:2439. doi: 10.1038/s41467-018-04823-3

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© The Author(s) 2018

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. 3 — Affinity purification of Y3IP1-HA. a ΔY3IP1/CGL59 mutant (LMJ.RY0402.195677) from the CLiP contains the paromomycin resistance cassette (CIB1) inserted at the 7th Exon (E7) of CGL59/Y3IP1 (Cre06.g280650). Exons, introns, and untranslated regions are shown as red boxes, black lines, and blue boxes, respectively. This strain also contains second CIB1 cassette in CAH2 locus (Cre04.g223050). The mutant was complemented with cDNA of Y3IP1 (c-Y3IP1) or Y3IP1-HA (c-Y3IP1-HA). b The growth of CLiP host strain as control, ΔY3IP1, and three c-Y3IP1 clones (2, 3, and 4) and c-Y3IP1-HA clones (1,3, and 9), under the photoautotrophic condition in the light of 50 μmol photons m⁻² s⁻¹. c Total cell proteins from control, ΔY3IP1, c-Y3IP1, and c-Y3IP1-HA strains were analyzed by immunoblotting using antibodies against Y3IP1, Ycf3, PsaA, and PSAL. The nitrocellulose membrane was stained with Ponceau for loading control. d The polypeptide composition of the affinity-purified Y3IP1-HA and Ycf3-HA. Polypeptides separated by SDS-PAGE were visualized by staining with Flamingo. e Immunoblotting of the purified Y3IP1-HA and Ycf3-HA with antibodies against PsaA and Ycf4