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. 2019 Jul 29;9:10984. doi: 10.1038/s41598-019-47387-y

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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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Knockout of α3 integrins in mSG-PAC1 cells inhibits mRNA expression of α6 integrins, but not the expression of E-Cadherin or Aqp5 mRNA in response to FGF2. Cells were incubated in medium containing EGF (black bars) or FGF2 (gray bars) (a) Expression of α3 mRNA in mSG-PAC1 cells cultured on tissue culture plastic in the presence of EGF or FGF2 for five days. Expression was normalized to β-actin and then plotted ± s.e.m. as the fold change in FGF2 normalized to expression in EGF. Data are from three independent experiments analyzed by Student’s T-Test. ***p < 0.001. (b) Expression of α3 mRNA in mSG-PAC1 and mSG-PAC1n (α3-deleted) cells cultured in a Matrigel/collagen I matrix in the presence of EGF for five days. Expression of α3 was normalized to β-actin and then plotted ± s.e.m. as the fold decrease in mSG-PAC1n compared to mSG-PAC1 cells. Data are from three independent experiments analyzed by Student’s T-test. ***p < 0.001. (c) Representative western blot probed for the α3 subunit in mSG-PAC1 and mSG-PAC1n cells cultured on tissue culture plastic for four days. The image of the full-length blot is provided in Supplementary Fig. S6. (d) Expression of α6 mRNA in mSG-PAC1 and mSG-PAC1n cells cultured as in panel B in the presence of EGF or FGF2 for five days. Data are from three independent experiments analyzed by two-way ANOVA. p < 0.05. Tukey’s post-hoc analysis was performed to correct for multiple comparisons. (e,f) Expression of E cadherin mRNA and Aqp5 mRNA in mSG-PAC1 and mSG-PAC1n cells cultured as described in panel D. Data are from three independent experiments analyzed by two-way ANOVA. Differences are not significant.