Fig. 10: Targeting KV2.1–CaV1.2 interactions reduces Mito Ca2+ and rescues neurotoxicity in NPC disease. | Nature Communications

Fig. 10: Targeting KV2.1–CaV1.2 interactions reduces Mito Ca2+ and rescues neurotoxicity in NPC disease.

From: NPC1-dependent alterations in KV2.1–CaV1.2 nanodomains drive neuronal death in models of Niemann-Pick Type C disease

Fig. 10

A Left, representative images of neurons transfected with ER-GCaMP6-150. Top right, ER-GCaMP6-150 localization. Bottom right, ER Ca2+ quantifications. N = 16 (CTL), n = 17 (U18), n = 4 (TTX) and n = 4 (TTX + U18) neurons were analyzed across 2 independent isolations. B Left, representative images of CTL (black), U18 (red), nifedipine (Nif) (blue) and Xestospongin C (XestoC) (green) neurons transfected with Mito-RCamP1h. Right top, Mito-RCamP1h localization. Right bottom, normalized Mito Ca2+. N = 35 (CTL), n = 33 (U18), n = 13 (Nif), n = 15 (Nif+U18), n = 8 (XestoC) and n = 9 (XestoC+U18) neurons were analyzed across 3 independent isolations. C Left, neurons expressing Mito-RCamP1h. Right top, Mito-RCamP1h localization. Right bottom, normalized Mito Ca2+. N = 13 (SCRBL), n = 14 (SCRBL + U18), n = 12 (CCAD) and n = 12 (CCAD + U18) neurons were analyzed across 2 independent isolations. D Left, representative images of neurons incubated with H2DCFDA. Right, quantification of ROS. N = 43 (SCRBL), n = 36 (SCRBL + U18), n = 39 (CCAD) and n = 64 (CCAD + U18) neurons were analyzed across 2 independent isolations. E Quantification of neuron mitochondrial membrane polarization. N = 28 (SCRBL; black), n = 48 (SCRBL + U18; red), n = 30 (CCAD; black) and n = 39 (CCAD + U18; purple) neurons were analyzed across 2 independent isolations. F Left, representative images of Live and dead channels from neurons in each condition. Quantification of neuronal viability. N = 394 (DMSO), 174 (U18), n = 94 (XestoC), n = 28 (XestoC + U18), n = 124 (SCRBL), n = 50 (SCRBL + U18), n = 85 (CCAD), n = 59 (CCAD + U18), n = 33 (Rosc) and n = 29 (Rosc + U18) neurons were analyzed across 5 independent isolations. All error bars represent SEM. Statistical significance was calculated using a two-way ANOVA test in (A)–(E) and One sample t test and Wilcoxon test in (F). ns: not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. ##P < 0.01; # indicates comparison with the CTL condition. SCRBL is scramble, U18 is U18666A, CCAD is calcium channel association domain, Rosc is Roscovitine, TTX is tetrodotoxin, Nif is Nifedipine, XestoC is Xestospongin C.

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