Copyright information:Taken from "P2X nucleotide receptors mediate caspase-8/9/3-dependent apoptosis in rat primary cortical neurons"Purinergic Signalling 2005;1(4):337-347.Published online Jan 2005PMCID:PMC2096553. (A) Rat PCNs were cultured on coverslips in 6-well plates and neurons were labeled with mouse NeuN monoclonal antibody () and cell nuclei with TO-PRO-3 (); merged picture shown in . (B) RNA was isolated from rPCNs maintained in culture for 7–10 days and RT-PCR was used to amplify mRNAs to P2XRs, 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 (). G3PDH primers were used to amplify G3PDH mRNA as a positive control. (C) P2XR protein expression was detected by Western blot analysis, as described in the Materials and methods. Human 1321N1 cells expressing the recombinant human P2XR or the empty expression vector pLXSN were used as positive (+) or negative (−) controls, respectively. Precision Plus Protein Standards are indicated as ‘75KD’ and ‘100KD.’