Secretomes of <i>Giardia</i> trophozoites and differentiated Caco-2 cells during interactions.

<p>(A) Number of secreted WB and GS proteins in axenic culture. (B) Number of secreted WB and GS proteins during host cell interactions. (C) Number of secreted Caco-2 cell proteins during interaction with <i>Giardia</i> WB and GS trophozoites. (D) and (E) GO term analyses of WB and GS secretomes during interaction with host cells. (F) and (G) GO term analyses of secreted Caco-2 cell proteins during interactions with <i>Giardia</i> WB and GS trophozoites.</p

The effects of ESPs on IEC cell signaling.

<p>(A) Western blot analyses of differentiated Caco-2 cells exposed to <i>G</i>. <i>intestinalis</i> ESPs through a Transwell insert for 2h or 6h in the absence of inflammatory stimuli (e.g. lipopolysaccharides or tumour necrosis factor-alpha). Blots were probed for total proteins (phosphorylated/non-phosphorylated forms) of mitogen activated proteins kinases (MAPKs, ERK1/2 and P38) and nuclear factor kappa beta (NFκB). (B) Western blot analyses after the induction of inflammation with LPS and TNF-alpha.</p

Gene expression changes in IECs after ESP exposure.

<p>(A) Heat-map of differentially expressed genes in the differentiated human colon carcinoma cell line Caco-2 exposed to <i>Giardia intestinalis</i> excretory secretory products. (B) Induction of AP-1 activity using a luciferase reporter plasmid transfected into Caco-2 cells. The cells were exposed to ESPs of <i>Giardia intestinalis</i> isolate WB or GS for 2h or 6h through a Transwell insert placed in the tissue culture plate (Mean ± SD, <i>n</i> = 4). (C) Relative RNA levels (Mean ± SD, <i>n</i> = 4) of inflammatory genes in differentiated Caco-2 cells incubated with different amounts of ESPs (5μg/ml and 10μg/ml) collected from <i>Giardia intestinalis</i> isolate WB or GS. Significant results in the panels are indicated by (*) for <i>P</i> < 0.05 or (**) for <i>P</i> < 0.01. (D) ELISA measurement (Mean ± SD, <i>n</i> = 3) of interleukin-8 (IL-8), chemokine (C-X-C motif) ligand 1 (CXCL1) and chemokine (C-C motif) ligand 20 (CCL20) in the medium of differentiated human colon carcinoma cell line, Caco-2, exposed to ESPs of <i>G</i>. <i>intestinalis</i> WB or GS isolate through a Transwell insert. No statistical significance was found between controls and treatments (<i>P</i> > 0.05).</p

Proteins secreted by differentiated Caco-2 cells specific to interaction with <i>Giardia intestinalis</i> isolates, WB and GS.

<p>Proteins secreted by differentiated Caco-2 cells specific to interaction with <i>Giardia intestinalis</i> isolates, WB and GS.</p

Top 50 identified proteins of secreted by differentiated Caco-2 cells co-incubated with <i>Giardia intestinalis</i> isolates, WB and GS <i>in vitro</i>.

<p>The proteins are ranked based on peptide score. Note that all listed proteins were also detected in Caco-2 cells culture without <i>Giardia</i>.</p

Top 50 identified proteins of <i>Giardia intestinalis</i> isolates, WB and GS, secreted into the medium of interaction with intestinal epithelial cells <i>in vitro</i>.

<p>The proteins are ranked based on their peptide score. Note that all listed proteins were also detected in axenic cultures.</p

Proteins secreted during <i>Giardia</i>-host cell interactions.

<p>During <i>Giardia</i>-host cell interactions <i>in vitro</i> specific proteins are released into the growth medium. The same types of proteins, mainly enzymes, are released from both the host and parasite (black) but there are also parasite-specific proteins being released (blue).</p

The binding and internalization of <i>Giardia intestinalis</i> excreted-secreted products (ESPs) to intestinal epithelial cells (IECs).

<p>Confocal microscopy images of differentiated human colon carcinoma cell line, Caco-2, incubated with Alexa Fluor488-labelled ESPs of <i>G</i>. <i>intestinalis</i> isolate WB or GS. Note that the colour of nucleus, stained with DAPI, was changed to red for better contrast.</p

Proteins specifically released by <i>Giardia intestinalis</i> isolates, WB and GS, during interaction with differentiated Caco-2 cells.

<p>Proteins specifically released by <i>Giardia intestinalis</i> isolates, WB and GS, during interaction with differentiated Caco-2 cells.</p

Selected groups of differentially expressed genes in differentiated human colon carcinoma Caco-2 cells exposed to excretory-secretory products of <i>Giardia intestinalis</i> isolates WB and GS via Transwell insert.

<p>Bold fonts indicate a significantly up-regulated gene whereas italic fonts indicate a significantly down-regulated gene.</p