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. 2019 Jul 3;7:118. doi: 10.3389/fcell.2019.00118

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Copyright © 2019 D’Souza, Blackburn, Kudlyk, Pokrovskaya and Lupashin.

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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Fluorescent Recovery After Photobleaching (FRAP) experiment reveals that an EELS is a stable organelle that exchanges its membrane and soluble content at a very slow rate. (A) Both soluble (mCherry-Lamp1) and membrane (Lamp2-GFP) cargo in EELSs HEK293T COG4 KO cells was simultaneously bleached using 488 nm and 546 nm lasers and recovery of fluorescent signal was analyzed at (B) 0 min, (C) 60 min, and (D) 120 min after the bleach. Superresolution images were collected using Zeiss LSM880 AiryScan microscope. Scale bars are 10 μm, white circles indicate the bleached region (E) The graph represents percentage of mCh (red) and GFP (green) signals over the course of 40 min. Error bars indicate SD of five fields that were imaged, 1–2 EELSs were chosen in every field.