QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 6. During encystation, ADI is translocated to the nuclei and inhibits CWP expression. (A) Results of western blotting that was performed after cytoplasm- and nuclear-fractionation assays using anti-ADI pAb at 24 and 48 hours post-encystation induction in wild-type (ADI) cells. L, cell lysate previous fractionation; C, cytoplasmic fraction; N, nuclear fraction (upper panel). Middle panel: anti-VSP1267 mAb is used to detect cytoplasm- or nuclear-fraction contamination. Lower panel: a representative time course of IFA shows ADI (red) distribution during encystation (ESVs in green). (B) Confocal microscopy and IFA using wild-type cells show that ADI (red) is translocated to the nuclei when the encysting cell is filled with ESVs (arrowheads). CWP1 is stained in green. (C) Top panels: representative differential interference contrast (DIC) and DAPI-staining images show a stable ADI-transgenic culture (Merge). Middle panels: direct IFA shows that trophozoites highly overexpressing ADI-HA (green) in the nuclei do not express CWP2 (red) during encystation (arrowheads in Merge). Bottom panels: closer analysis of three ADI-transgenic cells demonstrated that those expressing ADI-HA (green) in the nuclei (arrowheads in Merge) do not reveal CWP2 (red) expression after 24 hours of encystation, whereas those with low ADI-HA expression do (arrow in Merge). Scale bars: 10 µm.

Return to article