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. 2017 Apr 1;36(2):50–56. doi: 10.1089/mab.2017.0008

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© Satoshi Ogasawara et al., 2017; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.

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FIG. 1. — Specific detection of PODXL by PcMab-47 using flow cytometry. (A) CHO-K1 and CHO-K1/PODXL were treated with PcMab-47 (1 μg/mL; red) or control PBS (black) for 30 minutes at 4°C, followed by treatment with antimouse IgG-Oregon green. (B) LN229, LN229/PODXL, and PDIS-13 (LN229/PODXL-knockout cells) were treated with PcMab-47 (1 μg/mL; red) or control PBS (black) for 30 minutes at 4°C, followed by treatment with antimouse IgG-Oregon green. (C) Caco-2, MDA-MB-468, HEK-293T, and vascular endothelial cells were treated with PcMab-47 (1 μg/mL; red) or control PBS (black) for 30 minutes at 4°C, followed by treatment with antimouse IgG-Oregon green. (D) Lec1 (N-glycan-deficient CHO)/PODXL, Lec2 (sialic acid-deficient CHO)/PODXL, Lec8 (galactose-deficient CHO)/PODXL, and Lec13 (fucose-deficient CHO)/PODXL were treated with PcMab-47 (1 μg/mL; red) or control PBS (black) for 30 minutes at 4°C, followed by treatment with antimouse IgG-Oregon green. (E) PDIS-12 (N-glycan-deficient HEK-293T), PDIS-22 (sialic acid-deficient HEK-293T), PDIS-18 (galactose-deficient HEK-293T), and PDIS-20 (N-glycan/sialic acid/galactose-deficient HEK-293T) were treated with PcMab-47 (1 μg/mL; blue) or control PBS (black) for 30 minutes at 4°C, followed by treatment with antimouse IgG-Oregon green. Fluorescence data were collected using the Cell Analyzer EC800. PBS, phosphate buffered saline; PODXL, podocalyxin.