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Figure 5


Fig. 5. ADI expression increases during G. lamblia differentiation. (A) IFA and confocal microscopy show the G. lamblia encystation process. CWP2 (red) is synthesized and transported in ESVs in encysting trophozoites (ET, arrowhead). At the end of the encystation process, CWP2 is found in mature cyst walls (Cyst). Nuclei are stained with DAPI (blue). Scale bars: 10 µm. (B) Slot-blotting qualitatively shows gdh, adi and cwp2 gene expression at 0, 6, 24 and 48 hours of encystation in wild-type cells (WB/1267wt) and transgenic trophozoites (WB/1267-ADI+). The assay was performed in triplicate. (C) Analysis of gdh, adi and cwp2 gene expression by RT-PCR and optical density quantified by densiometry comparing both wild-type (WB/1267wt) and ADI-transgenic (WB/1267-ADI+) trophozoites. Data represent the means ± s.d. for n=4 of two independent experiments. (D) Western blotting using specific antibodies shows ADI, modified citrulline (Cit), VSP1267 and CWP2 protein expression in both wild-type (WB/1267wt) and ADI-transgenic (WB/1267-ADI+) trophozoites. Non-encysting trophozoites (NT, 10 µg) and 24-hour-encysting trophozoites (ET, 10 µg) were used for each sample.





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