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. 2023 Jan 13;30:4. doi: 10.1186/s12929-023-00898-3

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© The Author(s) 2023

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. 4 — Suppression of HDAC6 prevented the microtubule-p-ERK interaction. A Immunofluorescence staining for p-ERK (green), acetylated tubulin (red), α-tubulin (gray), and DNA (blue) in H460 cells treated with or without TSA (5 µM TSA) for 4 h. Box areas are enlarged. The head arrows indicate p-ERK puncta overlapping with α-tubulin. Colocalization of p-ERK with acetylated tubulin or α-tubulin was calculated as Manders' coefficient. The plot shows individual data and is presented as the mean ± SEM. *p < 0.05; ^#p < 0.05 (n = 10). Scale bar is 10 µm. B H460 and C A549 cells were treated with 5 µM TSA for 4 h. The lysate was separated into soluble (S) and pellet (P) fractions using a microtubule sedimentation assay and analyzed for p-ERK, ERK, acetylated tubulin, and α-tubulin by immunoblotting. The ratio of the pellet to the total fraction was calculated. Data are presented as the mean ± SEM. *p < 0.05 vs. control cells (n = 3). D H460 and E A549 cells were transfected with siRNA against HDAC6 (siHDAC6, 50 nM) or control siRNA (siCtrl). Cells were lysed, separated into soluble (S) and pellet (P) fractions using a microtubule sedimentation assay, and analyzed for p-ERK, ERK, acetylated tubulin, GRP78, and α-tubulin by immunoblotting. The ratio of the pellet to the total fraction was calculated. Data are presented as the mean ± SEM. *p < 0.05 vs. siCtrl cells (n = 3)