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. 2018 Sep 12;8:13713. doi: 10.1038/s41598-018-32054-5

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© The Author(s) 2018

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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Altered components of MAPK and AKT/mTOR signaling pahtways in HIF2α-overexpressing pancreata. (A) Periacinar cells positive for phosphorylated-ERK (pERK) are found in 2-week-old control pancreata but acinar cells are negative for pERK (white arrowheads, top panels). Increased pERK accumulation is observed in 2-week-old Pdx1-Cre;HIF2dPA pancreata (yellow arrowheads, bottom panels). Note that the increased expression was limited to areas with acinar-ductal metaplasia (yellow arrowheads, bottom panels). Acini with preserved morphology do not show extensive pERK accumulation (white arrowheads, bottom panels). pERK-positive stromal cells are also observed in Pdx1-Cre;HIF2dPA pancreata (asterisks, bottom panels). E-cadherin immunofluorescence was performed to visualize the pancreatic epithelium. (B) Double immunofluorescence analysis of phosphorylated RSP6 (pRSP6) and amylase in 2-week-old mice. pRPS6 (yellow arrows) is detected in amylase-positive acinar cells of control pancreata (top panels). In Pdx1-Cre;HIF2dPA pancreata, pRPS6 is detected in well-preserved acini (bottom panels, yellows arrows). However, areas with acinar-ductal metaplasia display decreased expression of pRPS6 (white arrows). (C) Double immunofluorescence analysis of pRSP6 and KRT19 in 2-week-old mice. Ductal cells (white arrows) are not positive for pRPS6 in both control pancreata (top panels) and Pdx1-Cre;HIF2dPA pancreata (bottom panels). (D) Clusterin is not expressed in control pancreas (top panels). Areas with acinar-ductal metaplasia in Pdx1-Cre;HIF2dPA pancreata display decreased expression of pRPS6 and activation of clusterin expression (white arrows). Yellow arrows indicate acini with normal morphology. Nuclei are stained with DAPI (blue). Scale bars = 50 µm.