Fig. 1.

Mdj1 localizes to mitochondrial nucleoid. (A,B) In vivo localization of Mdj1. Wild-type (wt) cells expressing Mdj1-GFP, Abf2-GFP or mt-GFP were analyzed by fluorescence microscopy. Images in the two right-most panels were overlaid (MERGE). Size bars (2 μm) are shown. (A) Cellular DNA was stained using DAPI. (B) Mitochondrial network was stained using MitoTracker. (C) Distribution of Mdj1 after centrifugation of mitochondrial lysates. Lysates prepared from isolated mitochondria were subjected to ultracentrifugation through a sucrose gradient. A portion of each fraction was analyzed for mtDNA content by PCR amplification of mtDNA ori5 fragment (mtDNA) and for protein content by immunoblot analysis using antibodies specific for Mdj1, Abf2, Ssc1 and Jac1.