Figure 1.

Purity of rat primary cortical neurons (rPCNs) and P2X₇R expression. (A) Rat PCNs were cultured on coverslips in 6-well plates and neurons were labeled with mouse NeuN monoclonal antibody (Panel a) and cell nuclei with TO-PRO-3 (Panel b); merged picture shown in Panel c. (B) RNA was isolated from rPCNs maintained in culture for 7–10 days and RT-PCR was used to amplify mRNAs to P2X_1–7Rs, as described in the Materials and methods. The amplified PCR products were resolved by gel electrophoresis, and data shown are representative of results from three independent experiments. Results are shown of PCRs performed in the presence (+) or absence (−) of reverse transcriptase (RT). G3PDH primers were used to amplify G3PDH mRNA as a positive control. (C) P2X₇R protein expression was detected by Western blot analysis, as described in the Materials and methods. Human 1321N1 cells expressing the recombinant human P2X₇R or the empty expression vector pLXSN were used as positive (+) or negative (−) controls, respectively. Precision Plus Protein Standards are indicated as ‘75KD’ and ‘100KD.’