ATP- and Bz-ATP-induced intracellular calcium fluxes in undifferentiated and neural-differentiated ESC. GlaserTalita La Banca de OliveiraSophia ChefferArquimedes BecoRenata MartinsPatrícia FornazariMaynara LameuClaudiana Miranda Costa JuniorHelio Coutinho-SilvaRobson UlrichHenning 2014 <p>ATP- and Bz-ATP-evoked P2X7R responses were determined by single-cell calcium imaging and microfluorimetry assays as described in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096281#s2" target="_blank">Methods</a>. (<b>A</b>) For calcium imaging, 10 µM of purinergic receptor agonists, ATP or Bz-ATP, were used to stimulate receptor-mediated intracellular calcium fluxes in undifferentiated and neural-differentiated cells (day 8). (<b>B</b>) For microfluorimetry-based calcium measurements, increasing concentrations of ATP or Bz-ATP were added and time kinetics of changes in fluorescence emission was recorded. Peak values of measured [Ca<sup>2+</sup>]<sub>i</sub> transients are reported. Nonlinear Regression was used for curve-fitting and calculating the EC50 values. (<b>C and D</b>) P2X7R inhibitors were studied regarding their effects in blocking ATP- and Bz-ATP-induced [Ca<sup>2+</sup>]<sub>i</sub> transients, as studied by calcium imaging. For these experiments, cells were pre-treated for 2 min with 10 µM KN-62 or 1 µM A438079 and then stimulated with 10 µM ATP or Bz-ATP. Data represent mean values ±S.E. of three independent experiments performed in triplicate.</p>