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. 2019 Sep 24;9:13778. doi: 10.1038/s41598-019-50360-4

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© The Author(s) 2019

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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Impact of Pepstatin A on neutrophil migration induced by patient gastric fluid samples set at pH 3 and pH 7.4. Pepstatin A (20 µg/ml) was added to gastric samples set at pH 3 and pH 7.4. Pepstatin A failed to significantly reduce the magnitude of neutrophil (PMN) migration across H292 lung epithelium induced by gastric samples set at pH 3 from patients off PPI therapy (A), but did significantly block neutrophil migration induced by gastric samples set at pH 3 from patients on PPI therapy (B). For gastric fluid samples set at pH 7.4, Pepstatin A failed to impact the magnitude of neutrophil migration induced by samples collected from patients off (C) and on (D) PPI therapy. A dilution of 1:4 in pH matched HBSS was used for all samples. Both on and off PPI therapy groups set at either pH 3 or 7.4 represent the mean +/− SEM of the average magnitude values (n = 3) of PMN migration by gastric fluid derived from 12 patients on PPI therapy and 12 patients off PPI therapy, diluted 1:4 in HBSS and treated with or without Pepstatin A (n = 12). Differences were considered significant between groups with and without Pepstatin A at p ≤ 0.05 and noted by the symbol (*). The p values were determined using an unpaired two-tailed student’s T test with equal variance within an internally controlled experiment. Panels A–D are representative internally controlled experiments conducted on at least two separate occasions yielding similar results.