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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/).

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Purification of RAVE–V₁and RAVE complexes.A, affinity purification via FLAG-tagged Rav2. Coomassie-stained SDS-PAGE of a peak fraction from affinity purification with FLAG-tagged Rav2 is compared with a peak fraction of V₁ affinity-purified via FLAG-tagged subunit G from a vma5Δ mutant. Positions of RAVE subunits are indicated in red, and V₁ subunits are indicated in black. Samples are from a single gel, cut for better labeling. B, gel filtration profile for a sample affinity-purified via FLAG-tagged Rav2 as described in Experimental procedures section. Below the profile is an SDS-PAGE of the indicated fractions. RAVE–V₁ is enriched in fraction 11, and RAVE is enriched in fraction 13. Fraction 12 appears to show a mixture of the two complexes. RAVE, Regulator of the H+-ATPase of Vacuoles and Endosomes.