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

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posted on 19.02.2013, 22:02 by Iulia I. Nita, Michal Hershfinkel, Daniel Fishman, Eyal Ozeri, Guy A. Rutter, Stefano L. Sensi, Daniel Khananshvili, Eli C. Lewis, Israel Sekler

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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