Figure 7: AP-2 ablation impairs the formation of axon initial segment.
From: Adaptor Protein 2 (AP-2) complex is essential for functional axogenesis in hippocampal neurons
(a) Representative images of cells transfected at 2 DIV with Nav1.6-GFP ± shRNA against AP-2 (AP-2KD). Neurons were fixed at 7 DIV and incubated with anti-GFP (green) and anti-ankyrin G (red). Arrow indicates AIS. (Inset) magnified images of AIS region to clearly visualize. (b) Mean value of the length of the AIS in each condition. Nav1.6-GFP only = 38.09 ± 1.75 μm (n = 24 cells), Nav1.6-GFP + AP-2KD = 3.61 ± 0.65 μm (n = 17 cells). ***p < 0.001. (c) Mean value of Pearson’s co-localization coefficient of Nav1.6-GFP and AnkyrinG in AIS and Soma. Control AIS = 0.58 ± 0.07, Control soma = 0.62 ± 0.11. AP-2KDAIS = 0.57 ± 0.14, AP-2KDsoma = 0.15 ± 0.59. (d–h) Measurement of Nav1.6-GFP at AIS and soma in control and AP-2KD neuron. (d,e) Dashed lines indicate line scan of Nav1.6-GFP distribution from AIS to Soma. (f and g) Profiles of line scan of Nav1.6-GFP in control (f) and AP-2KD (g) neurons. (h) Mean values of density of Nav1.6-GFP at AIS and soma in control and AP-2KD neurons. Control AIS = 513.63 ± 72.69 a.u., Control soma = 442.51 ± 76.19 a.u. (n = 21 cells). AP-2KDAIS = 119.43 ± 35.23 a.u., AP-2KDsoma = 1190.86 ± 145.63 a.u. (n = 15 cells).
