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. 2017 Aug 9;7:7642. doi: 10.1038/s41598-017-06588-z

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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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PI3K/AKT/mTOR signalling molecules associate with ΔF508 CFTR interactome. (a) CFTR was immunoprecipitated from WT HBE41o- and ΔF508 CFBE41o- cells and immunoblotting was performed for mTORC1/2 components, RICTOR, RAPTOR, mTOR, MAPKAP1 and GβL as well as CFTR and β-actin to confirm mass spectrometry observations (table left). A quantitative graph of (b) RICTOR and (c) MAPKAP1 in WT and ΔF508 CFTR immunoprecipitates. The results shown are representative of three independent experiments; the histograms represent the average and the error bars represent the standard deviation of the means. Statistical significance was examined using unpaired T-test analysis. Asterisk *represents p < 0.05. (d) Protein expression of mTOR, pmTOR Ser 2481, pAkt Ser 473 and total Akt (mTORC2) were determined by immunoblotting in WT HBE41o- and ΔF508 CFBE41o- cell lysates in addition to CFTR and β-actin. (e) Protein expression of mTOR, pmTOR Ser 2448, p70 S6 kinase Thr 389 and total 70 S6 kinase (mTORC1) were determined by immunoblotting in WT HBE41o- and ΔF508 CFBE41o- cell lysates in addition to CFTR and β-actin. (f) A quantitative graph of mTOR from WT HBE41o- and ΔF508 CFBE41o- cell lysates, the results shown are representative of three independent experiments; the histograms represent the average and the error bars represent the standard deviation of the means. Statistical significance was tested by unpaired T-test analysis. Asterisk* represents p < 0.05 (g). Protein expression of pAkt Ser 473, p70 S6 kinase Thr 389, Akt and p70 S6 kinase were determined by immunoblotting in ΔF508 CFBE41o- (37 °C) and ΔF508 CFBE41o- cells temperature shifted to 27 °C in addition to CFTR and β-actin.