FIG. 2.

Expression of IRS mRNAs and proteins in wild-type brown adipocytes during differentiation. (A) Quantitative real-time PCR by the ABI Taqman procedure was used to measure the transcript numbers for each IRS in total RNA extracted from wild-type cells on days 0, 3, and 6 of differentiation. Details of RNA preparation, primers, probes, and the method of quantification are described in Materials and Methods. The data are presented as relative numbers of transcripts per cell using calibration curves prepared for each IRS. (B) Protein expression of IRS-1 and IRS-2 in wild-type brown adipocyte differentiation. Specific antibodies against IRS-1 or IRS-2 were used in Western blot analysis as described in Materials and Methods. (C) Blow-up graph of panel A to better illustrate the expression levels of IRS-3 and IRS-4 mRNAs during differentiation. The results are means from two to four experiments.