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. 2012 May 14;14(8):1299–1317. doi: 10.1111/j.1462-5822.2012.01800.x

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Copyright © 2012 Blackwell Publishing Ltd

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Fig. 4 — Expression and localization of tetracycline-inducible EhCP-A5-HA. A. Immunoblot analysis of the expression of tetracycline-inducible EhCP-A5-HA. The E. histolytica transformant expressing tetracycline-inducible EhCP-A5-HA or the control transformant was cultured in the presence of 1 or 5 µg ml⁻¹ tetracycline for the indicated times. The lysates were analysed by immunoblot analysis with the anti-HA antibody. B. Localization of tetracycline-inducible EhCP-A5-HA. The E. histolytica transformant expressing tetracycline-inducible EhCP-A5-HA was labelled with LysoTracker Red, further cultured with 5 µg ml⁻¹ tetracycline for the indicated times, and subjected to IFA with the anti-HA antibody. Bars, 10 µm. C. Localization of EhCP-A5-HA and CPBF1. The E. histolytica line that coexpressed CPBF1-myc constitutively and EhCP-A5-HA in a tetracycline-inducible manner was labelled with LysoTracker Red, further cultured with 5 µg ml⁻¹ tetracycline for the indicated times, and subjected to immunoblot analysis (upper panel) or immunofluorescence assay with the rabbit polyclonal anti-HA and mouse monoclonal anti-myc antibodies (lower panel). Magnified images of the area of interest showing EhCP-A5-HA being accumulated within the vesicles that did not colocalize with CPBF1 are shown in rectangles. Bars, 10 µm.