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. 2020 May 22;10:8558. doi: 10.1038/s41598-020-65353-x

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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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HE4-mediated STAT3 activation and upregulation of IL8 and HIF1A is blocked by STAT3 inhibition (A) Upregulation of phospho-STAT3 in SKOV3 cells, HUVECs, and PBMCs by rHE4 treatment. Upregulation of HIF1α by rHE4 observed in HUVECs and PBMC. STAT3 activation and HIF1A levels were ablated by treatment with a STAT3 inhibitor. Boxes separate images from the same gel for each cell type. Blots were either subsequently reprobed for various antibodies, or stripped and reprobed in the case of STAT3/p-STAT3. (B) ELISA revealed increased secretion of IL8 by PBMC treated with 20 nM rHE4 from 4–24 h post-treatment, which was suppressed by STAT3 inhibition. (C) ELISA revealed increased secretion of IL8 by PBMC treated with 1 nM rHE4 from 4–24 h post-treatment, which was suppressed by STAT3 inhibition. (D) ELISA revealed IL8 secretion by PBMC was increased as early as 2 h post-treatment with rHE4. Error bars represent standard deviation of ≥3 biological replicates. *p < 0.05, **p < 0.005, ***p < 0.005, ****p < 0.0005.