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. 2013 Apr 23;2(3):525–539. doi: 10.1002/mbo3.88

Figure 1.

Figure 1

Ubiquitination is a dynamic process in Giardia. (A) Schematic representation of the in vitro encystation process. Trophozoites were induced to encyst and after 48 h cyst were obtained. Principal organelles are indicated as N: nucleus, VD: ventral disc, F: flagella, ESV: encystation-specific vesicles, and CW: cyst wall. (B) Immunodetection of Ub and Ub-protein conjugates. Trophozoites were induced to encyst, and cells were collected at different times during the process (0, 6, 12, 24, and 48 h). Extracts of different encystation stages were subjected to SDS-PAGE and immunoblotted with monoclonal anti-Ub. Immunoblotting detection of CWP1 (26 kDa) was used as the encystation marker. (C) Immunolocalization of Ub and Ub-protein conjugates. Cells were collected, fixed with p-formaldehyde, and used for immunofluorescence analysis. Ub and Ub-conjugated proteins were detected with anti-gUb. Anti-CWP1-TAMRA was used to detect CWP1 protein. Nuclei were stained with DAPI. The different stages are indicated as follows: (0 h/T) trophozoites, (6 h) encysted cell 6 h postinduction, (24 h) encysted cell 24 h postinduction, and (48 h/C) cyst obtained after 48 h of induction. Full arrows indicate the peripheral spots. Open arrows indicate encystation-specific vesicles (ESVs). Scale bar is 10 μmol/L. DIC is differential interference contrast microscopy.