Extended Data Fig. 9: Axon initial segment structural plasticity occurs in SCN2A+/− neurons and is rescued by CRISPRa.
From: CRISPR activation for SCN2A-related neurodevelopmental disorders
a. Representative images of SCN2A+/+ (black) and SCN2A+/− (cyan) human stem-cell-derived neurons immunostained with antibodies against ankyrin-G (green) and MAP2 (magenta). Arrowheads denote start and end points used to quantify AIS length. b. Quantification of AIS length. SCN2A+/+: n = 56 cells, 3 dishes. SCN2A+/−: n = 56 cells, 3 dishes. ****p < 0.0001. Mann-Whitney test. c. Representative images of SCN2A+/− neurons expressing Scn2a-rAAV-CRISPRa-mCherry (purple) and mCherry-negative internal SCN2A+/− controls (cyan). Immunostaining against ankyrin-G and MAP2. d. Quantification of AIS length. SCN2A+/−: n = 122 cells, 3 dishes. SCN2A+/− + CRISPRa: n = 23 cells, 3 dishes. ****p < 0.0001. Mann-Whitney test. Circles are individual AIS calculations. Box plots are medians, quartiles, and 100% tails.
