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. 2016 May 20;5(5):e001625. doi: 10.1161/JAHA.114.001625

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© 2016 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.

This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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Ubiquitylation of TRPM4. A, GST pull‐down experiment using GST‐tagged S5a (S), a subunit proteasome that recognizes ubiquitylated proteins, and GST alone (G) to pull down TRPM4 from HEK293 cells transiently transfected with WT TRPM4, A432T, and A432T/G582S individually. The input fraction is shown at left, and the pull‐down fraction is shown at right. B, Quantification of each fraction and comparison of WT to variant ratios between fractions. Quantification was done on 3 different Western blots. GST indicates glutathione‐S‐transferase; n.s., not significant; WT, wild type.