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Fig. 5. Nd1-L mRNA associates with TLS. (A) Gel mobility shift assay using radioactive-labeled Nd1-L 3'-UTR probe. In vitro synthesized 493 nucleotide RNA probe for Nd1-L mRNA was radioactively labeled with ³²P. The ³²P-labeled probe was incubated with TLS protein prepared from rabbit reticulocytes and the complexes were resolved by native gel electrophoresis. A 50-fold molar excess of cold probe inhibited the binding of the radioactive probe to TLS. From left to right; lane 1, probe only; lane 2, probe with no competitor; lanes 3, 4, and 5, fivefold, 50-fold and 500-fold molar excess, respectively, of cold probe was added to the binding reaction together with the radioactive-labeled probes. (B) 100-fold molar excess of unlabeled irrelevant transcripts (205 nt transcript encoding pcDNA3.1/Myc-His polylinker site; 780 nt transcript encoding ß-actin 3'-UTR containing a zip code sequence) were challenged to in vitro binding of TLS and ³²P-labeled Nd1-L probe.

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