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. 2021 Apr 22;296:100703. doi: 10.1016/j.jbc.2021.100703

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© 2021 The Authors

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

PMC Copyright notice

Pulldowns of MBP-tagged subunit C and Vph1NT with RAVE and RAVE–V₁.A, gel filtration fractions containing RAVE–V₁ or RAVE were incubated with MBP-C and anti-FLAG M2 beads as indicated, then transferred to a column for elution. All blots were loaded with input (I), flowthrough (F), wash (W), and fractions collected after addition of FLAG peptide to elute bound protein. MBP-C was also incubated with beads in the absence of FLAG-tagged RAVE as a control. B, RAVE–V₁ was incubated with MBP-Vph1NT as described in A in the presence and absence of MBP-C as indicated. MBP-Vph1NT with no added RAVE–V₁ is included as a control. MBP, maltose-binding protein; MBP-C, MBP-tagged subunit C; RAVE, Regulator of the H⁺-ATPase of Vacuoles and Endosomes; Vph1NT, N-terminal domain of V_o subunit Vph1.