VSMCs contain CCR8 mRNA and membrane-associated CCR8 antigen. (A) RNA blot analysis of CCR8 mRNA in VSMCs. Aliquots (10 μg) of total RNA from Jurkat cells (Ju) or from confluent human VSMCs (SMC) were size-fractionated on agarose gels and hybridized to ³²P-labeled human CCR8 cDNA as described in “Materials and methods.” GAPDH is shown as a control for loading of samples. The location of 28 and 19S markers is indicated. The cDNA was generated as detailed in “Materials and methods,” and the identity of the fragment as CCR8 mRNA was confirmed by sequence analysis. (B) Immunoblot analysis of CCR8 in VSMCs. Human VSMC membranes were prepared as detailed in “Materials and methods,” and the solubilized membrane protein and CCR8 expression was analyzed by immunoblot analysis using a polyclonal anti-CCR8, which identified an approximately 40-kDa band.