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. 2021 Jan 13;22(2):753. doi: 10.3390/ijms22020753

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Flow cytometric analyses of EpCAM+/CD44v6+/ALDH+/CD133+ cells in SW948-FR cells isolated from SW948-FR/subcutaneous (SQ) tumors. Alexa fluor-tagged antibodies at the indicated laser lines were used to isolate: (A) EpCAM+/ECadherin- cells; (B) CD44v6+/ALDH1+ cells from EpCAM+/ECadherin-cells; (C) CD133+ cells from CD44v6+/ALDH1+ cells. (D) Immunofluorescence staining shows colocalization of CD44v6 (Red) and YB-1 (Green) in EpCAM+/CD44v6+/ALDH1+/CD133+ (CICs), scale bar, 50 μm. (E) Percentages of EpCAM+/E-cadherin-, CD44v6+/ALDH1+, and CD44v6+/ALDH1+/CD133+ on sorted cells were assessed by flow cytometry on freshly purified CRC cells isolated from subcutaneous (SQ) SW948-FR tumor cells. (F) Western blots are shown for EpCAM+/CD44v6-/ALDH-/CD133- (Non-CICs) and EpCAM+/CD44v6+/ALDH+/CD133+ (CICs) by probing with E-Cadherin, EpCAM, CD44v6, ALDH1, YB-1, and CD133 antibodies. FACS, immunofluorescence and WB data are representative of three experiments. Enrichment of CICs in FR cells compared to sensitive pairs in Figure 2E are presented as mean ± SD (n = 3); *, p < 0.01. Student’s t test was used to assess the significance. The experiment was performed three times and representative data are shown.