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. 2018 May 22;9(6):596. doi: 10.1038/s41419-018-0624-8

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Western blot analysis reveals all four p38 MAPK isoforms (α, β, γ and δ) are expressed in wild type and SOD1^G93A motor neuron lysates. Mouse brain lysate was loaded as an antibody control for p38 MAPKβ. The upper band is only detected in spinal cord extracts and may represent a posttranslational modified form of p38 MAPKβ. b Lentiviral delivery of shRNA targeted against p38 MAPKα reduces its levels in primary motor neurons. Transduced motor neurons express GFP. c Speed profiles of H_CT-containing signalling endosomes in wild-type motor neurons transduced with scrambled shRNA (black squares), SOD1^G93A motor neurons transduced with scrambled shRNA (red diamonds) and SOD1^G93A motor neurons transduced with p38 MAPKα shRNA (blue triangles). Axonal transport has been quantified only in GFP-positive motor neurons. Comparison of the curves reveals that knocking down p38 MAPKα in SOD1^G93A motor neurons (blue triangles) accelerates axonal transport to wild-type speeds (black squares) (wild type + scrambled shRNA: 89 carriers, 17 axons; SOD1^G93A + scrambled shRNA: 104 carriers, 24 axons; SOD1^G93A + p38 MAPKα shRNA: 99 carriers, 20 axons; 4 independent experiments). d Lentiviral delivery of scrambled shRNA has no effect on levels of p38 MAPKα in primary motor neurons. Transduced motor neurons express GFP. e Speed profiles of H_CT carriers in untreated SOD1^G93A motor neurons (green circles) and SOD1^G93A motor neurons transduced with scrambled shRNA (scram; red diamonds). Comparison of the curves reveals that the scrambled shRNA construct (red diamonds) has no effect on axonal transport speeds (SOD1^G93A untreated: 45 carriers, 7 axons; SOD1^G93A + scrambled shRNA: 104 carriers, 24 axons; 4 independent experiments). f Speed profiles of H_CT carriers in untreated wild-type motor neurons (green circles) and wild-type motor neurons transduced with scrambled shRNA (scram; black squares). Comparison of the curves reveals that scrambled shRNA constructs have no effect on axonal transport speeds (wild-type untreated: 62 carriers, 5 axons; wild type + scrambled shRNA: 89 carriers, 17 axons; 3 independent experiments)