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. 2012 Apr 24;287(28):23852–23863. doi: 10.1074/jbc.M111.328708

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Subcellular fractionation of heart homogenates. A, TAG content in the fractions obtained by sucrose density gradient centrifugation of heart homogenates of wild-type and Plin5^−/− mice under fed and fasted conditions. Open bar, wild-type mice, and filled bar, Plin5^−/− mice. B, immunoblotting of Plin5 and Plin2 in the fractions of A. For fed animals, results of long exposure of the blots are also shown. Arrowhead, Plin5 and Plin2. Asterisk, nonspecific band, which was not recognized by another lot of anti-Plin5 antibody (Progen) raised against the same carboxyl-terminal epitope. C, protein contents in the same fractions. D, distribution of markers of mitochondria (cytochrome c oxidase subunit 3 (COX3)), endoplasmic reticulum (stearoyl-CoA desaturase 1 (SCD1)), and cytosol (glyceraldehyde-3-phosphate dehydrogenase (GAPDH)), and actin filament through the gradients. E, TAG content in the fractions after centrifugation through a wider sucrose-density gradient. Heart homogenates from fed wild-type (open bar) and Plin5^−/− (filled bar) mice were analyzed. Open circle, specific gravity (Sp. gr.) of each fraction. F, distribution of Plin5, Plin2, and marker proteins in the same fractions as in E. Calnexin was used as a marker of endoplasmic reticulum.