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. 2019 Jul 1;30(14):1716–1728. doi: 10.1091/mbc.E18-12-0811

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© 2019 Gravotta et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 8: — AP-1 controls apical biosynthetic delivery of HA-mMeg-GFP. (A) Scheme of controlled trypsin cleavage assay for biosynthetic surface delivery. MDCK cells stably expressing ER-retained FM4-HA-mMegalin-GFP are incubated at 37°C with D/D solubilizer, and with trypsin, added apically or basolaterally, to induce ER export and subsequent cleavage of HA-mMegalin-GFP upon cell surface arrival. (B) Western blotting detection with an anti-GFP antibody of cleaved ∼130 kDa HA-mMegalin-GFP. (C) Quantification of the results. Values derived from three independent experiments. *p value < 0.05. Note that single γ1 and combined γ1/γ2 knockdown but not single γ2 knockdown cause missorting of HA-mMeg-GFP.