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. 2017 Jul 3;7:4546. doi: 10.1038/s41598-017-04874-4

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© The Author(s) 2017

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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HDAC6 inhibition in PAH-PASMCs causes Bax-induced cell death by increasing acetylation of cytosolic Ku70. (A) Representative Western blots of acetyl(K539)-Ku70 and total Ku70 in PAH-PASMCs after treatments with two HDAC6 inhibitors Tubastatin A and ACY-775 or siHDAC6 for 48 hours. Data were expressed as the ratio of acetyl(K539)-Ku70/total Ku70. (B) Expression of acetylated(K539)-Ku70 and Ku70 was assessed in distal PAs from control (n = 4) and PAH patients (n = 5) by Western blot. Data were expressed as the ratio of acetyl(K539)-Ku70/total Ku70. (C) Apoptosis was measured using Annexin V staining (green) in control (n = 3 cell lines) and PAH-PASMCs (n = 3 cell lines) pre-treated or not with Bax inhibitory peptides (BIP-V5, 200 μM) or control peptides before being exposed to indicated HDAC6 inhibitors for 48 hours. Representative immunofluorescence images of apoptotic Annexin-V-positive cells (green) stained for DAPI (blue) and corresponding quantification are shown. Scale bar = 50 μm. Protein expression was normalized by Amido black (AB). ***P < 0.001 and ****P < 0.001.