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. 2021 Jun 29;5:61. doi: 10.1038/s41698-021-00192-1

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© The Author(s) 2021

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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Fig. 3 — a PTX3 expression in whole cell lysates (WCL) and conditioned media (CM) of pancreatic stellate cell line (PS1), cancer cell lines (MiaPaCa2, Patu-T, Patu-S, Capan1, AsPc1) by Western blot analysis and b densitometric quantification (n = 4). HSC70 was used as a loading control. c Western blot depicting expression of PTX3 in WCL and CM of PS1 cells 72 h after transfection with 20 nM of PTX3 siRNA (siRNA) or non-targeting siRNA (NT), with HSC70 loading control and densitometric quantification (n = 3). Summary data are expressed as mean with standard error mean. d Duration of PTX3 silencing resulting from siRNA transfection was assessed. Western blot of PTX3 expression in WCL and CM of pancreatic stellate cells (PS1) from 24–168 h. e Representative H&E and immunofluorescence images of mini-organotypic cultures of cancer (MiaPaCa2) and stellate (PS1) cells transfected with PTX3 siRNA or NT siRNA, demonstrating HABP expression. Scale bar = 100 μm. Summary quantification data from mini-organotypic co-cultures of cancer (MiaPaCa2) and stellate (PS1) cells transfected with PTX3 siRNA or NT siRNA. f Quantification of HABP expression in images (n = 6, 8 images/gel). g Percentage of invading cells presented as the mean percentage relative to NT control. h Cell proliferation assessed by the measurement of the cell layer thickness. Summary quantification data from mini-organotypic monocultures of stellate (PS1) cells transfected with PTX3 siRNA or NT siRNA. i Quantification of HABP expression in images (n = 6, 8 images/gel). j Percentage of invading cells presented as the mean percentage relative to NT control. k Cell proliferation assessed by the measurement of the cell layer thickness. Sections from three experimental replicates were analyzed for mini-organotypics, resulting in 27 high-power field measurements (n = 9, 3 sections/gel). OT1, OT2, OT3: organotypic biological repeats. b Kruskal–Wallis test; WCL, CM: ****P < 0.0001, c Mann–Whitney U test, **P < 0.01,***P < 0.001, f–k Wilcoxon matched-pairs test, *P < 0.05, **P < 0.01, ***P < 0.001. Summary data are expressed as median and interquartile range.