Effect of NCLX expression or activity on glucose dependent cytosolic calcium responses.

<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>

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