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Taken together, these results showed that the GS/H7-induced structural and functional brush border injuries are both adhesion-dependent. Our findings also demonstrate that GS/H7-induced changes in the distribution and activity of brush border-associated SI hydrolase and GLUT5 transporter are dependent on the GS/H7-induced disassembly of the F-actin cytoskeleton. Previous reports have shown that decreased TER results from alterations of the TJ-associated proteins, ZO-1 and claudin-1, after Giardia infection of cell monolayers (Buret et?al., 2002; http://www.selleckchem.com/GSK-3.html Scott et?al., 2002) and in human biopsy specimens from patients with chronic giardiasis (Troeger et?al., 2007). We confirmed that a decrease in TER develops in Caco-2/TC7 cell monolayers infected with G.?intestinalis strains GS/H7, WB/C6 or WB/1267 (moi?=?8, 24?h of infection) (Table?2). Time-course monitoring of TER in G.?intestinalis GS/H7-infected Caco-2/TC7 cell monolayers showed a time-dependent decrease of TER starting at 12?h p.i. (control: 774?��?29; GS/H7 3?h p.i. 771?��?32; GS/H7 12?h p.i. 585?��?45; GS/H7 24?h p.i. 350?��?23?��?cm2). We conducted an in-depth analysis of the distribution of http://www.selleckchem.com/screening/tyrosine-kinase-inhibitor-library.html two functional TJ-associated proteins: occludin (Furuse et?al., 1993) and claudin-1 (Furuse et?al., 1998) in G.?intestinalis-infected Caco-2/TC7 cell monolayers (moi?=?8, 24?h of infection). Indirect immunolabelling of occludin and CLMS analysis showed that the protein was localized at http://en.wikipedia.org/wiki/VAV2 cell-to-cell contact points in uninfected Caco-2/TC7 cells, and the typical honeycomb-like pattern can be observed (Fig.?7A). In contrast, apical infection of Caco-2/TC7 cell monolayers with G.?intestinalis strains GS/H7, WB/C6 or WB/1267 induced modification of the cell-to-cell organization characterized by an obvious loss of the typical honeycomb structure (Fig.?7A). Occludin distribution was modified, now showing a brightly stained continuous band lining each cell-to-cell contact. Phosphorylated and non-phosphorylated forms of occludin have both been described (Farshori and Kachar, 1999). It has been established that non-phosphorylated occludin has a cytoplasmic localization, and that it is easily extracted using Triton X-100. In contrast, phosphorylated occludin, located at cell-to-cell contact points, is resistant to extraction with Triton X-100. Occludin distribution was analysed by Western blotting of Triton X-100-soluble and Triton X-100-resistant fractions of G.?intestinalis GS/H7-infected Caco-2/TC7 cells. In Triton X-100-soluble fractions (Fig.?7B), a 65?kDa band corresponding to the non-phosphorylated form of occludin was observed. A densitometric analysis of the Triton X-100-soluble fraction of G.?intestinalis GS/H7-infected cells revealed that non-phosphorylated occludin was significantly lower (P?