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. 2018 Jan 11;8:436. doi: 10.1038/s41598-017-18884-9

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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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ABT-263 reduces endotoxin-induced mucous cell hyperplasia in vivo in a Bik-dependent manner. (A) Experimental outline for testing therapeutic efficacy of ABT-263 in LPS-induced MCH in mice. (B) Representative micrographs of lung tissue sections stained with Alcian-Blue (AB) and H&E from LPS-challenged mice treated with vehicle or ABT-263 (2 mg/Kg) are shown. Quantification of mucous cell numbers per mm BL. (C) Representative micrographs showing activated (cleaved) caspase 3 or Ac-Casp3 (green) among Scgb1a1-positive (red) secretory cells in mouse axial airways. The relative fold-change in the number of ac-Casp3+ secretory cells in LPS-challenged mice treated with vehicle or ABT-263. (D) Representative micrographs showing TUNEL-positivity (green) in Scgb1a1+ (red) secretory cells in mouse axial airways treated with ABT-263 and DAPI-stained nuclei (blue). The relative fold-change in the number of TUNEL+ secretory cells in mice challenged with LPS and treated with vehicle or ABT-263. (E) STAT-1 phosphorylation in HAECs following 0, 15, and 60 minutes of IL-13 treatment. Cropped Western blots are displayed. (F) Bik and Bcl-2 mRNA levels in IL-13 treated STAT1 ^−/− and STAT1 ^+/+ MEFs compared with the respective non-treated cells. (G) Immunoprecipitation with anti-Bcl-2 antibodies of proteins extracted from HAECs treated with nothing or ABT-263. Bik levels are reduced in the pull-down while increased in the flow-through of HAECs treated with ABT-263 compared with non-treated controls. Cropped Western blots are displayed. (H) Representative micrographs of lung tissue sections from LPS-challenged mice stained with AB-H&E and quantification of mucous cell numbers per mm BL in mice treated with vehicle or ABT-263 (2 mg/Kg) following LPS challenge. Scale = 20 µM; Data shown as mean ± SEM (n = 5–10 mice/group); *p < 0.05; **p < 0.01; ***p < 0.001.