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. 2018 Feb 1;137(1):53–67. doi: 10.1007/s11120-017-0477-5

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© The Author(s) 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.

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Fig. 7 — Subcellular localization of the YL2 protein. a Subcellular localization of the YL2 protein was determined by GFP assay. 35S::GFP (upper panel) and 35S::YL2-GFP (lower panel) were transiently expressed in rice protoplast cells. Left to right, GFP fluorescence, chlorophyll autofluorescence, merged images, and bright field images. Bar: 5 μm. b Suborganelle localization of the YL2 protein in the chloroplast. Intact chloroplasts (Chl) were isolated from WT leaves then separated into thylakoid membrane (Th), envelope (En), and stroma (St) fractions. Proteins were separated by 10% SDS-PAGE, electroblotted onto PVDF membranes then probed with polyclonal antisera against D1 (thylakoid marker), RbcL (stroma marker), Tic110 (envelope marker), and YL2. c YL2 represents an intrinsic protein. Thylakoid membrane proteins isolated from WT plants were incubated with 1 M NaCl, 200 mM Na₂CO₃, and 6 M urea for 30 min at 4 °C. PsbO (extrinsic membrane protein) and CP47 (intrinsic membrane protein) were used as markers. Membranes without salt treatment were used as a control (CK)