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Fig. 1 | Molecular Brain

Fig. 1

From: Lysosomal iron modulates NMDA receptor-mediated excitation via small GTPase, Dexras1

Fig. 1

Liable iron is available in CA1 and is released in an activity dependent manner. a. Following application of the iron sensitive dye Calcein-AM, CA1 had the largest response to PIH application when compared to DG and CA1 (n = 5). Top 2 panels show pseudocolor images of hippocampal slices dyed with Calcein-AM dye both before (left) and after 100 μM PIH (Right; an average of 15 s). Purple indicates the biggest change in fluorescence. Bottom left shows the change in fluorescence over time normalized to baseline fluorescence, and the bar graph is an average of the change in fluorescence in CA1 and DG (*p < 0.05). b. Live cell imaging was performed with primary hippocampal neurons using fluorescence-based iron de-quenching imaging method with Calcein-AM dye (n = 7). Depolarization of the neurons with 50 mM KCl induces a 77 % decrease in fluorescence. When PIH is added to the culture, fluorescence returns to 89 % of control (p < 0.001). In contrast there is no decrease in fluorescence in Dexras1 KO hippocampal cultures when PIH is applied. c. Summary of live cell imaging with inhibitors which block nNOS and DMT1. Primary hippocampal neurons are pretreated with inhibitors for 30 min and an imaging experiment was performed as shown in Fig B. Inhibitor treatment significantly reduced the iron released into the cytosol (p < 0.05). d. Approximately 2,000 primary hippocampal neurons were seeded in 96 well plates and labeled with PhennGreen SK dye (20 μM). The fluorescent signal was measured by BioTek multiplate reader linked with automated reagent dispenser for KCl (a final concentration of 50 mM) (n = 3). The y-axis is arbitrary fluorescence change

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