P2Y₁₂ Receptor on the Verge of a Neuroinflammatory Breakdown

Figure 1

P2Y₁₂ antibodies validation and RT-PCR analysis. (a) Scheme of human P2RY₁₂ gene location, transcript variants [29, 30], and protein structure, with amino acid epitopes recognized by the used antibodies (Table 1) and highlighted in color (intra1, intra2, and intra fl, red circle; c-ter, green oval). Species conservation for each epitope was calculated by using BLAT tool of UCSC genome browser [31]. (b) Total protein extracts from SH-SY5Y or HEK293 cells expressing Myc-tagged P2Y₁₂ receptor were subjected to Western blot analysis with the indicated antibodies. (c) Total protein extracted from human, rat and mouse brain, from primary mouse microglia (mMG) and rat oligodendrocyte (OL) cultures were subjected to Western blot analysis with the indicated antibodies. For intra2 antibodies,  lots AN01/02/04/0502/0602 were used. (d) RT-PCR using primers specific for P2Y₁₂ mRNA was performed on total RNA from rat microglia (rMG) and OL. Control lanes show RT-PCR performed without reverse transcriptase enzyme.