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. 2018 Oct 4;115(43):E10099–E10108. doi: 10.1073/pnas.1814552115

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Copyright © 2018 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 4. — Procollagen/Sec23/LC3 puncta colocalize with ubiquitination machinery. (A) Airyscan slice showing TagBFP2-LC3 and GFP-proα2(I)^G610C puncta colocalization with Halo-Sec23 and Apple-CUL3, an E3 ubiquitin ligase. All large FP-CUL3 puncta were confirmed to be colocalized with FP-LC3 (Bottom Left image of the whole cell) and procollagen (zoomed panels, Right), and some also contained FP-Sec23 (zoomed panels, outline). (B) Airyscan slice showing colocalization of TagBFP2-LC3 puncta with Halo-KLHL12 adaptor for CUL3 ubiquitin ligase (Bottom Left) and with GFP-proα2^G610C(I) and Apple-Sec23 (zoomed panels, outlines). All Halo-KLHL12 puncta colocalized with GFP-proα2^G610C(I) were confirmed to be also colocalized with TagBFP2-LC3 (top images of the whole cell). (C) Airyscan slice showing colocalization of Halo-LC3 puncta with Apple-Keap1 adaptor for CUL3 ubiquitin ligase (Bottom Left) and with GFP-proα2^G610C(I) and TagBFP2-Sec23 (zoomed panels, outlines). In all zoomed panels, yellow outlines of LC3-positive puncta are projected in white onto other channels. Individual blue channels are displayed in cyan. [Scale bars: 10 µm (whole cell) and 2 µm (zoom).]