Fig. 2: Loss of NPC1 function alters voltage-dependent CaV1 channel distribution.
A Left: Representative super-resolution Airyscan images taken at a focal plane near the plasma membrane (PM) from CTL (black) and U18-treated (red) neurons fixed and immunolabeled for CaV1.2. Right, quantification of PM CaV1.2 total intensity, cluster density and cluster size of CTL (black) and U18-treated (red) neurons in the soma (left, yellow) and dendrite (right, orange) regions. N = 46–48 (CTL) and n = 58–59 (U18) neurons, and n = 145 (CTL) and n = 161 (U18) dendrites were analyzed across 5 independent isolations. B Left, representative super-resolution TIRF localization maps of CTL (black) and U18-treated (red) neurons fixed and immunolabeled for CaV1.2. Right, quantification of PM CaV1.2 cluster density, cluster size, and nearest cluster distance of CTL (black) and U18-treated (red) neurons in the soma region. N = 20 (CTL) and n = 21 (U18) neurons were analyzed across 3 independent isolations. C Same as (A) only neurons fixed and immunolabeled for CaV1.3. N = 32 (CTL) and n = 38 (U18) neurons, and n = 104 (CTL) and n = 123 (U18) dendrites were analyzed across 3 independent isolations. D Same as B, only immunolabeled for CaV1.3. N = 16 (CTL) and n = 18 (U18) neurons were analyzed across 2 independent isolations. All error bars represent SEM. Statistical significance was calculated using Mann–Whitney (two-tailed) and Unpaired t tests (two-tailed) in (A)–(D). ns: not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. CTL is control and U18 is U18666A.
