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. 2004 Aug;78(15):7958–7968. doi: 10.1128/JVI.78.15.7958-7968.2004

FIG. 3.

FIG. 3.

Core protein cofractionates with mitochondria. (A) Western blot analysis of HM and LM fractions isolated from core protein-expressing HeLa and Jurkat Tet-Off cells by differential centrifugation. Cell equivalents from each fraction were loaded, and the distributions of FLAG-tagged core protein, mitochondrial heat shock protein 70 (mtHsp70), and the ER-resident calreticulin protein were analyzed. (B) Western blot analysis of HM and LM fractions of two Huh-7 cell lines (21-5 and 20-1) expressing the full-length replicon with core protein- and organelle-specific antibodies. (C) Jurkat homogenates were further separated by sucrose density gradient centrifugation to obtain purified mitochondria and MAMs. Microsomes and cytosolic fractions were prepared by differential centrifugation of the same cell homogenates. Equal protein amounts (30 μg) from each fraction were analyzed by Western blotting with antibodies specific for FLAG-tagged core protein, cytochrome c (Cyto c), calreticulin, and heat shock protein 90α (Hsp90α).