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

A. 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. Silencing NCLX expression inhibits glucose-induced Ca2+ entry in primary β cells. Representative fluorescent traces of cytosolic Ca2+ 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 Fig. 2A. Insert. Shows representative images of MIN6 cells co-transfected with the Dharmacon siGLO Red transfection reagent. The scale bar is 10 µm. C. NCLX dominant negative construct inhibits glucose dependent cytosolic Ca2+ 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. D. Averaged rates of cytosolic Ca2+ responses of Fig. 4B, n = 10 (*P<0.05). E. Averaged rates of cytosolic Ca2+ responses of Fig. 4C, n = 10 (*P<0.05). F. Averaged cytosolic Ca2+ amplitudes of Fig. 4B, n = 10 (*P<0.05). G. Averaged cytosolic Ca2+ amplitudes of Fig. 4C, n = 10 (*P<0.05).

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