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. 2017 Sep 22;7:12207. doi: 10.1038/s41598-017-12367-7

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© The Author(s) 2017

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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Basal characteristics of SQ20B cells, SQ20B-CSCs and FaDu cells. (A) EGFR (180 kDa), HER2 (184 kDa) and HER3 (189 kDa) basal expression in SQ20B cells, SQ20B-CSCs and FaDu cells. Protein expression was analysed with WES™ (a simple western blotting technique using an automated capillary-based size sorting system) and expressed graphically as the GAPDH ratio. (B) Phospho-AKT (Ser473) (63 kDa) and phospho-MEK1/2 (Ser217/221) (52 kDa) basal expression in SQ20B cells, SQ20B-CSCs and FaDu cells. Protein expression was analysed with WES™, and expressed graphically as the GAPDH ratio (protein-of-interest (POI)/GAPDH). (C) Microscopic observation (63×) of SQ20B cells stained with phalloidin (green actin) and DAPI (blue nucleus). (D) Microscopic observation (63×) of SQ20B-CSCs stained with phalloidin (green actin) and DAPI (blue nucleus). (E) Microscopic observation (63×) of a SQ20B cell stained with phospho-EGFR-FITC (green) and DAPI (blue nucleus). (F) Microscopic observation (63×) of a SQ20B-CSC stained with phospho-EGFR-FITC (green vacuoles) and DAPI (blue nucleus). Statistically significant differences are expressed as *p < 0.05, **p < 0.01 and ***p < 0.001. Each experiment was performed in triplicate.