PAT domain protein gene expression in Adfp+/+ (+/+) and Adfp−/− (−/−) mice. (A) Northern blot analysis. A total of 20 μg of total RNA isolated from WAT, BAT, muscle, and liver was separated on a 1% agarose gel; the RNAs were transferred to a nylon membrane and probed with cDNA of perilipin (PlinA), Tip47, S3-12, and Adfp. (B) Western blot analysis. A total of 20 μg of total protein extracts isolated from WAT, BAT, muscle, and liver was separated by polyacrylamide gel electrophoresis (PAGE) using a 4 to 15% gradient gel; the proteins were transferred to a nylon membrane and detected using antisera raised against perilipin and Tip47 and enhanced chemiluminescence kits.