N-terminal amino acid alignment and relative expression of Tim50a and Tim50. (A) Compared to Tim50, Tim50a contains additional sequence that contains a putative nuclear localization sequence (NLS) upstream of the translational start of Tim50. (B) Schematic representation (not to scale) of Tim50a and Tim50 showing the locations of the primers used in the PCR reactions. (C) The relative expression of Tim50 versus Tim50a in four cancer cell lines. (Upper panel) A standard PCR reaction using cDNA from each cell line as template and ComFor+ComRev primers yields a product that could be from Tim50 and Tim50a. Water serves as a negative control while Tim50a plasmid cDNA serves as a positive control. Nested PCR (lower panel) demonstrates that Tim50a is detected in two cell lines but accounts for only a small fraction of the products from the TIMM50 gene. The cDNA from the following cell lines was used: A-549 (non-small cell lung cancer), NIH: OVCAR-3 (ovarian cancer), MDA-MB-231 (breast cancer) and HCT-15 (colon cancer). The lower band in the NIH: OVCAR-3 nested PCR reaction is a non-specific product.
Xu et al. BMC Cell Biology 2005 6:29 doi:10.1186/1471-2121-6-29