Figure 3.
Expression of leptin mRNA and other adipose-specific mRNAs during adipose development from s.c. implanted 3T3-F442A preadipocytes. The level of expression leptin mRNA and mRNAs encoding other adipocyte markers was determined by Northern blot analysis of total RNA isolated from fat pads derived from the implanted preadipocytes described in Fig. 2. (A) Northern blots of RNA isolated from adipose tissue derived from implanted 3T3-F442A preadipocytes, from epididymal white adipose tissue (eWAT), and from in vitro differentiated 3T3-F442A adipocytes. Approximately 10 μg of total RNA was loaded per lane. The blot was hybridized to a 32P-labeled probe for leptin and then stripped and reused for hybridization to probes for C/EBPα, SCD1, 422/aP2, and 28S rRNA. (B) Plots of the results in A showing the relative levels of each transcript expressed as percentage of the level of transcript in RNA from epididymal adipose tissue. Hybridization signals on Northern blots (shown in A) were quantified using a phosphorimager and were normalized to the corresponding signals from the rRNA. Transcript levels are expressed as percentage of the mean transcript level of eWAT. Results are represented as the mean ± range (n = 2).