Extended Data Fig. 5: Manipulation of the cytoskeleton causes changes in axonal pearling.

a. Western blots and quantification showing the efficiency of shRNA-mediated knock down (KD) of ßII spectrin magenta shows ßII spectrin and yellow shows GAPDH. Scr, scramble. b. Example micrographs of axons from neurons treated with indicated drugs. Scale bar: 200 nm. c. Plots showing dimensions of NSVs (left) and connectors (right) from axons in (a). Mean and SEM are shown. N = 3 independent cultures n = 100 axons each. n.s., not significant. d. Additional example micrographs of axons from neurons infected with lentivirus carrying scramble shRNA or shRNA against ßII spectrin. Scale bar: 200 nm. e. Super Plots of Fig. 3d, showing experimental variability N = 3 independent cultures and n = 100 axons each. f. Additional example micrographs of axons from neurons treated with indicated drugs. Note that these neurons are treated with DMSO or Latrunculin A for 1 hour. Scale bar: 200 nm. g. Super Plots, showing experimental variability of Fig. 3a. N = 1 and n = 100 axons. h. Additional example micrographs of axons from neurons treated with indicated drugs. Scale bar: 200 nm. i. Super Plots of Fig. 3f, showing experimental variability. N = 3 independent cultures and n = 100 axons each. j. Additional example micrographs of axons from neurons treated with indicated drugs. Scale bar: 200 nm. k. Super Plots, showing experimental variability from experiments in Extended Data Fig. 5j. N = 3 independent cultures and n = 100 axons each. In all super plots median and 95% confidence intervals are shown. Each color represents one replicate. Each dot is one axon. All data analyzed using Kruskal-Wallis test, followed by Dunn’s multiple comparison test.

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