Effect of NCLX expression or activity on glucose dependent cytosolic calcium responses. Iulia I. Nita Michal Hershfinkel Daniel Fishman Eyal Ozeri Guy A. Rutter Stefano L. Sensi Daniel Khananshvili Eli C. Lewis Israel Sekler 10.1371/journal.pone.0046649.g004 https://plos.figshare.com/articles/figure/_Effect_of_NCLX_expression_or_activity_on_glucose_dependent_cytosolic_calcium_responses_/232788 <p><b>A.</b> Real time PCR analysis of mRNA NCLX expression normalized to GAPDH in pancreatic primary β cells transfected with siNCLX vs. siControl, n = 3 (*P<0.05). <b>B.</b> Silencing NCLX expression inhibits glucose-induced Ca<sup>2+</sup> entry in primary β cells. Representative fluorescent traces of cytosolic Ca<sup>2+</sup> in pancreatic primary β cells transfected with either siNCLX or siControl loaded with Fura 2 AM and stimulated with high glucose following the same experimental paradigm described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046649#pone-0046649-g002" target="_blank">Fig. 2A</a>. <b>Insert.</b> Shows representative images of MIN6 cells co-transfected with the Dharmacon siGLO Red transfection reagent. The scale bar is 10 µm. <b>C.</b> NCLX dominant negative construct inhibits glucose dependent cytosolic Ca<sup>2+</sup> changes in primary β cells. Representative fluorescent traces of primary β cells transfected with dnNCLX or control vector (pcDNA) loaded with Fura 2 AM and treated with high glucose when indicated. <b>D.</b> Averaged rates of cytosolic Ca<sup>2+</sup> responses of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046649#pone-0046649-g004" target="_blank">Fig. 4B</a>, n = 10 (*P<0.05). <b>E.</b> Averaged rates of cytosolic Ca<sup>2+</sup> responses of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046649#pone-0046649-g004" target="_blank">Fig. 4C</a>, n = 10 (*P<0.05). <b>F.</b> Averaged cytosolic Ca<sup>2+</sup> amplitudes of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046649#pone-0046649-g004" target="_blank">Fig. 4B</a>, n = 10 (*P<0.05). <b>G.</b> Averaged cytosolic Ca<sup>2+</sup> amplitudes of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046649#pone-0046649-g004" target="_blank">Fig. 4C</a>, n = 10 (*P<0.05).</p> 2013-02-19 22:02:49 nclx glucose cytosolic calcium