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. 2019 Dec 18;9:19369. doi: 10.1038/s41598-019-55867-4

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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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Regulation of GM2 expression in CSC-like cells by MAPK pathway. (a) FACS analysis of GM2 expression in MIA PaCa-2 cultured in 3D conditions with or without 1 μM PD0325901 treatment. Mean fluorescence intensities (MFIs) are shown on the right side. (b) Phase contrast images of MIA PaCa-2 cultured in 3D conditions with or without 1 μM PD0325901 treatment. (c) Immunoblotting for TGF-β1 signaling in sphere cells with or without 1 μM PD0325901 treatment. Seven days after sphere formation, sphere cells were cultured for an additional 2 days with or without 10 ng/ml TGF-β1. The blot images were cropped to highlight the pSmad2, pSmad3, Smad2/3, TGFβRI, TGFβRII and β-actin bands using the anti-pSmad2, anti-pSmad3, anti-Smad2/3, anti-TGFβRI, anti-TGFβRII and anti-β-actin antibodies. The full blot images are presented in the Supplementary western blot dataset. (d) Matrigel invasion assays performed in sphere cells. **p < 0.01.