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. 2020 Jul 25;13:105. doi: 10.1186/s13041-020-00644-y

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — HDAC6 is required for dendrite development in hippocampal neurons. a Knockdown effect of HDAC6 shRNA on HDAC6 expression. The HDAC6 level (red) was decreased in HDAC6 shRNA expressing neurons (venus), but expression of HDAC6 shRNA resistant HDAC6 mutant recovered the HDAC6 level in HDAC6 knockdown neurons. Scale bar, 20 μm. b Hippocampal neurons expressing HDAC6 shRNA. Scale bar, 50 μm. c Sholl graphs of neurons expressing Venus or HDAC6 shRNA or HDAC6 shRNA together with shRNA resistant HDAC6. Data represents average of three independent experiments (n = 10 for each group in each experiment). d-f Quantification of the number of primary dendrites (d), longest dendrite length (e) and total dendrite length (f). g, h HDAC6 did not affect axon specification. The number of axons in neuron was calculated by AnkG positive neurite. g Hippocampal neurons expressing Venus (green) or both Venus and HDAC6 shRNA were stained by AnkG (red). Nucleus was stained by Hoechst (blue). Scale bar, 50 μm. h Quantification of the percentage of neurons with 0, 1, 2, or more axons. (control, n = 62; HDAC6 KD, n = 64) *p < 0.05, **p < 0.01 by two-tailed Student’s t-test (unless otherwise indicated). The symbol # represents the number. Error bar means ± S.E.M.