Figure 7

P2Y ₁ receptor localizes in lipid rafts. Cerebella were removed from P21 rats and cell protein extracted in lysis buffer for 30 min on ice. Supernatants obtained after a centrifugation for 10 min at 15000 × g were loaded on SDS-PAGE (100 μg/lane), transferred to nitrocellulose and probed with anti-P2Y₁ receptor antiserum, in the absence (-) or presence (+) of the immunogenic peptide (A). Lipid rafts-enriched fractions were then prepared from total cerebellar tissue (B) or from purified cerebellar synaptosomes (C), both obtained from P21 rats. P2Y₁ receptor is recovered in rafts fractions 3 (B), in the absence of the immunogenic peptide, or 4–7 (C), together with the selective lipid rafts markers flotillin-2 (B, C) or GM1 (C), but not the transferrin receptor (C), a protein known to be excluded from lipid microdomains.