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. 2019 Aug 27;9:69. doi: 10.1186/s13578-019-0333-8

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© The Author(s) 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 1 — Effect of 4-PBA on secretion of rFVII-160R and rFVIIwt. Cells stably expressing rFVII-160R or rFVIIwt were treated with 0 (untreated), 5, 7.5 or 10 mM 4-PBA for 48 h. a–d FVII:Ag levels in conditioned medium (secreted) or cell lysates (intracellular) were examined by ELISA. To adjust for variations in cell number, the FVII:Ag levels were adjusted to the corresponding total protein content in the respective wells. a FVII:Ag levels in conditioned medium or cell lysates from untreated cells expressing rFVIIwt (black bars) or rFVII-160R (white bars). Values are reported as mean ± SD from at least three independent experiments performed in duplicates (unpaired t-test, ****p ≤ 0.0001). b Ratio between secreted/intracellular FVII:Ag levels in untreated cells. Values are reported as mean ± SD from at least three independent experiments performed in duplicates (unpaired t-test, ****p ≤ 0.0001). c, d FVII:Ag levels in the conditioned medium or cell lysates from cells expressing rFVII-160R (c) or rFVIIwt (d), treated with 4-PBA were examined by ELISA, and a ratio between secreted/intracellular FVII:Ag was calculated. Values are expressed relative to untreated cells and are reported as mean ± SD from at least three independent experiments performed in duplicates (one-way ANOVA, ****p ≤ 0.0001, ***p ≤ 0.001, **p ≤ 0.01). F7 mRNA levels from cells expressing rFVII-160R (e) or rFVIIwt (f), treated with 4-PBA were analysed by Taqman qRT-PCR. Quantitation was performed using the comparative CT method with 18S as the endogenous control gene and untreated cells as the calibrator. Bars represent mean fold-change in expression compared to untreated cells. Values are reported as mean ± SD from at least three individual experiments with four biological parallels (one-way ANOVA ****p ≤ 0.0001)