Fig. 2. Expression of P2Y1 mRNA and P2Y1 protein. RT-PCR analysis of subtypes of ATP receptors (A) in the primary culture of subepithelial fibroblasts and (B) in 18Co cells. (C) Western blotting analysis of P2Y1 receptors. Samples were loaded as follows. Lane A: subepithelial fibroblasts cultured in medium containing 10% FCS (control) (20 µg per lane). Lane B: subepithelial fibroblasts incubated in medium with 1 mM dBcAMP without FCS for 2 hours (20 µg per lane). Lane C: 18Co cells (50 µg per lane). Lane D: 3-week-old rat cerebrum (100 µg per lane). Samples were run on 10% SDS-polyacrylamide gels, transferred onto blotting membranes and detected with anti-P2Y1 antibody.