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. 2019 Jul 23;9:10684. doi: 10.1038/s41598-019-47116-5

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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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CD133/CD44 expressing cells are more in 3D and SMG. Dot plot for the surface expression of cancer stemness markers CD44 and CD133 analysed with FACS. CD44 stained with FITC, along Y axis and CD133 stained with PE along X axis (A). The experimental panel are grouped as rows (no experimental platform 1^st row, 3D 2^nd row and SMG 3^rd row) and treatment groups AI and AE in columns (no treatment 1^st column, Bafilomycin treatment (AI) 2^nd column and Rapamycin treatment (AE) 3^rd column) the top left pane represents control (untreated). The red gating boxed represent unique CD133/CD44 high cells in SMG. The graph representing cells positive for CD44 expression (quadrants 1 + 2) (B), cells positive for CD133 expression (quadrants 2 + 3) (C) and dual positive population counted form quadrant 2 (D). The experiment was repeated thrice and the graphs are represented as mean ± S.D. Significance measured using unpaired t test with Welch’s correction of individual groups with control is represented over the bar and the comparison between AI /AE with 3D/SMG is marked separately. The black bars represent control, light bars represent 3D and dark bars SMG. The bars with (/) lines represent AI and bars with (\) lines represent AE.