Research Article

Iodothyronine Interactions with the System L1 Amino Acid Exchanger in 3T3-L1 Adipocytes

Figure 7

LAT1, a catalytic subunit for System L1 transport, is expressed in 3T3-L1 adipocytes at the mRNA level but LAT2 is not. Reverse transcriptase PCR (RT-PCR) using gene-specific primers designed towards either LAT1 or LAT2 was performed with template cDNA produced from RNA extracted from 3T3-L1 adipocytes. Samples were separated using 2% agarose alongside a 1kb molecular weight marker (Roche Applied Science). (a) LAT1 mRNA is expressed in 3T3-L1 adipocytes. Following agarose gel electrophoresis, a band of 150 bp was observed corresponding to the expected PCR amplicon using LAT1-specific primers as detailed in the text. (b) LAT2 mRNA is not expressed in 3T3-L1 adipocytes. Following agarose gel electrophoresis, no PCR amplicon was detected when 3T3-L1-derived cDNA was used as a template. However, PCR using LAT2-specfic primers and cDNA derived from rat skeletal muscle did amplify a band of the expected size (465 bp), as shown, which demonstrates the functionality of the primers used.
726098.fig.007a
(a)
726098.fig.007b
(b)