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. 2020 Jul 29;39(36):5876–5887. doi: 10.1038/s41388-020-01399-5

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© The Author(s) 2020

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. 3 — a Western blot analyses for pAKT (S473, serine 473; T308, threonine 308), AKT, pERK, and ERK in control (shCTL) and stable PIERCE1 KD (shP#1 and shP#3) A549 cell lines. HSP90 was used as a loading control. Western blot analyses for pAKT (S473) and AKT in control and stable PIERCE1 KD A549 cell lines 36 hours after transient transfection of PIERCE1-GFP (b) and in time dependent TGFβ-treatment conditions (c). GFP was used to detect overexpressed PIERCE1. ACTIN was used as a loading control. Western blot analyses for pAKT (S473), AKT under TGFβ and TNFα treatment conditions during transient transfection of PIERCE1 (d) or doxycycline-induced PIERCE1 overexpression condition (e) in A549 cells. Doxycycline (1 μg/mL) was administered for 48 hours. ACTIN was used as a loading control. Overexpression of PIERCE1 was verified using GFP or PIERCE1 antibodies.