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MICU1 and MICU2 stabilize each other's expression and interact with MCU.

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posted on 2013-02-07, 00:50 authored by Molly Plovanich, Roman L. Bogorad, Yasemin Sancak, Kimberli J. Kamer, Laura Strittmatter, Andrew A. Li, Hany S. Girgis, Satya Kuchimanchi, Jack De Groot, Lauren Speciner, Nathan Taneja, Jonathan OShea, Victor Koteliansky, Vamsi K. Mootha

A. Whole cell lysates from HEK293T cells stably expressing a control shRNA (shGFP and shLACZ) or a shRNA targeting MICU1 (shMICU1_a and shMICU1_b) or MICU2 (shMICU2_a) were analyzed using qPCR and western blot. The relative mRNA is reported using β-actin as an endogenous control and normalized to shGFP for each target. Whole cell lysates were blotted with anti-MICU1, anti-MICU2 and control anti-ATP5A. B. Whole cell lysates from HEK293T cells stably expressing FLAG-GFP or FLAG-MICU1 were lysed and blotted with anti-MICU2, anti-FLAG and control anti-ATP5A. C–D. Mitochondria isolated from HEK293T cells stably expressing MCU-FLAG (C) or FLAG-MICU1 (D) were solubilized with 0.2% DDM and subjected to anti-FLAG immunoprecipitation. Immunoprecipitates and lysate were blotted with anti-FLAG, anti-MICU1, anti-MICU2 and control anti-ATP5B and anti-SDHB.