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. 2009 Oct 29;284(52):36491–36499. doi: 10.1074/jbc.M109.064527

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Ablation of Tb-Nbp35, Tb-Cfd1, and Tb-Nfs1 causes accumulation of non-thio-modified cytosolic tRNAs but does not influence mitochondrial import. A, RNA isolated from total cell (tot) and digitonin-extracted mitochondrial pellets (mt) (5) of uninduced (-Tet) and induced (+Tet) Tb-Nbp35-RNAi cell line were separated on APM-containing polyacrylamide gels that allow to separate thio-modified (thio-tRNA) from non-thio-modified tRNA (non-thio-tRNA). The top panel shows the ethidium bromide-stained gel. The bottom panel shows the corresponding Northern blot probed for the indicated tRNAs. The numbers at the bottom indicate the percentage (±S.E.) of non-thio-modified cytosolic tRNA before (-Tet) and after (+Tet) induction of RNAi for 72 h. The graph compares the extent of mitochondrial localization of the non-thio-modified tRNAs in uninduced cells (-Tet, set to 100%) to the one in induced cells (+Tet). Standard errors are indicated. B, same analysis as in A but for the Tb-Cfd1 RNAi cell line. C, same analysis as in A and B but for the Tb-Nfs1 RNAi cell line.