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. 2011 Dec 28;6(12):e29584. doi: 10.1371/journal.pone.0029584

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Pantakani et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Sub-cellular fractionation was performed with NSC34 cell extract. The cytosolic fraction (S-100) and the membrane fraction (P-100) were analyzed by immunoblot with ZFYVE27 antibody. Asterisks denote the non-specific band. The monomeric ZFYVE27 (mo-ZFYVE27) and SDS-resistant dimeric ZFYVE27 (di-ZFYVE27) are indicated. The membrane fraction (P-100) was subjected to Triton X-114 phase separation and equal volumes of aqueous phase (peripheral) and detergent phase (integral) fractions were analyzed by immunoblotting. The blot was stripped and re-probed with COX4 antibody, a marker for the integral membrane protein.