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. 2023 Apr 14;299(6):104718. doi: 10.1016/j.jbc.2023.104718

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© 2023 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

VPS33B and VPS16B form a high molecular weight complex.A, immunoblot analysis of HEK293 and DAMI cell lysates subjected to BN-PAGE detected native protein complexes (arrowhead) containing VPS33B and VPS16B that comigrated with 480 kDa marker. B, SDS-PAGE immunoblot showing co-IPs with anti-FLAG antibody of lysates from HEK293 cells cotransfected with Myc-VPS16B or Myc-VPS33B, plus VPS33B-3xFLAG or VPS16B-3xFLAG, respectively, probed with anti-Myc antibody. C, BN-PAGE immunoblots of co-IP experiments using anti-FLAG antibody and lysates from HEK293 cells cotransfected with Myc-VPS16B plus VPS33B-3xFLAG or Myc-VPS33B plus VPS16B-3xFLAG. Blots show Myc-tagged subunit co-IPs with FLAG-tagged subunit, indicating consistent formation of VPS33B–VPS16B complex comigrating with 480 kDa marker (arrowhead). BN-PAGE, blue native polyacrylamide gel electrophoresis; co-IP, coimmunoprecipitation; DAMI, megakaryoblastic DAMI cell line; HEK293, human embryonic kidney 293 cell line; VPS, vacuolar protein sorting protein.