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. 2021 Mar 8;118(11):e2019046118. doi: 10.1073/pnas.2019046118

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Copyright © 2021 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 2. — Ca_v1.2α1 in crude mitochondria is an artifact of Ca_v1.2α1 in the ER. (A) Representative immunoblots of brain mitochondria ultrapurification show depletion of Ca_v1.2α1 and MAM immunoreactivity from ultrapure mitochondria. Subcellular compartments are color coded as cytosol (white), PM (teal), ER (blue), MAM (pink), OMM (light gray), IMM (black), and mitochondrial matrix (dark gray) (n = 4, combined from three experiments). (B) Z score of marker enrichment relative levels in ultrapure mitochondria fraction during mitochondria ultrapurification. Highest enrichment fractions are boldly outlined. (C) Immunoprecipitation of Ca_v1.2α1 enriches Ca_v1.2α1 relative to IP input (Upper). (D) Representative immunoblots show bands from PNGase F treatment following Ca_v1.2α1 antibody immunoprecipitation from whole-cell homogenates and semipure mitochondria to evaluate for the presence of N-glycosylation protein modifications (n = 4, combined from two experiments).