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. 2021 Nov 8;221(1):e202103156. doi: 10.1083/jcb.202103156

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© 2021 Zheng et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S1. — Identification and validation of PEX19 as a MARCH5-interacting protein. (A) Reproducibility of the proximity labeling experiments to identify MARCH5-interacting proteins. Pearson’s correlation coefficients between replicate (Rep.) MS results for PUP-IT^MARCH5 and control samples. (B) PEX19 peptides containing GGE modification on lysine residues identified by MS. Red highlights the modification sites. Blue highlights the peptides in the context of full-length PEX19. (C) Multiple MARCH5-interacting candidates, including PEX19, are modified with biotin-Pup, showing higher-molecular-weight bands on WBs. HeLa cells were cotransfected with MARCH5-PafA-Myc and V5-tagged interacting candidate proteins. Proteins labeled by biotin-Pup were enriched by streptavidin magnetic beads, then analyzed by WB with anti-V5 antibodies. Black arrow, unmodified PEX19-V5 (expected, 35 kD); red arrows, biotin-Pup–modified PEX19-V5 (expected, 55 kD for monomodification and 75 kD for dimodification).