Skip to main content
. Author manuscript; available in PMC: 2021 Mar 29.
Published in final edited form as: Nat Cell Biol. 2020 Sep 21;22(10):1239–1251. doi: 10.1038/s41556-020-00577-7

Extended Data Fig. 7. The acetylcholinesterase transcript shows ribosome drop-off and defective production of protein at the NMJ in SMA.

Extended Data Fig. 7

a, Translation efficiency of Acetylcholinesterase from brain at early and late stage and in spinal cord at late stage, and after SMN-restoring Antisense Oligonucleotides treatment (11). The average ± SEM from n=3 biologically replicates in n=2 technical replicates is shown. For control brain early stage the average ± SEM from n=3 biologically replicates in n=3 technical replicate is shown. Translation efficiencies were obtained as the ratio of changes in polysomal and total RNA normalized to Tuba4a. Significant differences were determined using a two-sided t- test. b, Luciferase assays were performed in NSC34 expressing high or low levels of SMN in n=9 biologically independent replicates. c, Luciferase assays for testing the contribution of the first five codons of AChE with respect to four alanines. In (b) and (c) the number of biologically independent experiments is reported. Results are the average ± SEM. Significant changes were assessed using one-sided t-test. d, Brain, spinal cord and muscle lysates of early-symptomatic SMA mice were compared to controls using western blot. Experiments were repeated independently 3 times with similar results. e, Representative images for control (left) and early-symptomatic SMA mouse (right) neuromuscular endplates. Acetylcholine receptors (AChR) were labelled using alpha-bungarotoxin (BTX) and acetylcholinesterase (AChE) using fasciculin-2 (FCC). Top panels show FCC / BTX overlap, middle and bottom panels individual channels in greyscale. Endplates n=65 from control and n=73 endplates from SMA mice from 6 muscles and 3 mice/genotype were imaged and analysed. Scale bar: 10 μm. f, FCC and BTX average intensity were determined for 2 FDB muscles in 3 control and 3 SMA mice. The number of endplates per mouse is reported. Results are the average fluorescence intensity ± SEM. Significant differences were determined using a two-sided t-test. Statistical source data and unprocessed blots are provided in Source data Extended data Fig. 7.