Figure 7. Role of ankyrin B in the regulation of the periodic lattice structure.

(A–B) DIV 10 neurons from ankyrin B knockout (KO) mice were immunostained for βII spectrin and a dendritic marker Map2 and imaged. (A) Conventional image of βII spectrin and Map2. (B) 3D STORM of βII spectrin. The image is taken from the green-boxed region of the neuron in Figure 7—figure supplement 1. The enlarged STORM image and autocorrelation analyses of boxed regions are shown in (B1) and (B2). Similar results were found in four independent experiments. (C) Conventional βII spectrin image from wild-type (control) and ankyrin B KO DIV 10 neurons. The fluorescence intensity is coded by color, with red indicating higher expression. (D) The relative fluorescence intensity of βII spectrin in dendrites and axons of wild-type and ankyrin B KO neurons (n = 14 neurons for wild-type and n = 13 neurons for ankyrin B KO; three independent experiments).

DOI: http://dx.doi.org/10.7554/eLife.04581.020

Figure 7—figure supplement 1. The βII spectrin structure in axons of ankyrin B KO neurons.

(A) Reconstructed image of an ankyrin B KO DIV 10 neuron. βII spectrin was subjected for 3D STORM image in both dendrites and axons. The STORM image of dendrites in green-boxed region is shown in Figure 7. The STORM image of axon in white-boxed region is shown in (B). (C) Enlarged STORM image of the boxed region in (B). (D–E) Fourier transform and autocorrelation analyses of the axon segment shown in C.

DOI: http://dx.doi.org/10.7554/eLife.04581.021

Figure 7—figure supplement 2. Formation of the periodic lattice structure in dendrites of ankyrin B knockout neurons depends on βII spectrin.

(A–B) Ankyrin B knockout neurons were immunostained for adducin (green) and MAP2 (magenta), and imaged by conventional (A) and 3D STORM (B) microscopy. Magnified STORM images of adducin for arrow-indicated regions are shown in (B-1) and (B-2). (C–D) Ankyrin B knockout neurons were infected with βII spectrin-shRNA expressing adenovirus, immunostained for adducin and MAP2, and imaged by conventional (C) and 3D STORM (D) microscopy. Infected neurons were marked by GFP signal expressed from the virus (not shown). Magnified STORM images of adducin from arrow-indicated region are shown in (D-1) and (D-2). (E) Average autocorrelation analysis of adducin distribution in dendrites of ankyrin B knockout neurons (n = 9 neurons, four independent experiments). (F) Average autocorrelation analysis of adducin distribution in dendrites of ankyrin B knockout neurons treated with βII spectrin-shRNA expressing adenovirus (n = 15 neurons, three independent experiments).

DOI: http://dx.doi.org/10.7554/eLife.04581.022