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Fig. 1 | Journal of Biomedical Science

Fig. 1

From: Upregulation of MUC5AC production and deposition of LEWIS determinants by HELICOBACTER PYLORI facilitate gastric tissue colonization and the maintenance of infection

Fig. 1

MUC5AC and Lewis X/Y production by guinea pig gastric epithelial cells. MUC5AC and Lewix X/Y was evaluated on the basis of the intensity of fluorescence of guinea pig primary gastric epithelial cells or gastric tissue specimens stained with mouse anti-MUC5AC or anti-LeX/anti-LeY antibodies (Ab) and secondary antibodies conjugated with fluoresceine isothiocyanate (FITC) and counterstained with DAPI or phalloidine. a- guinea pig primary gastric epithelial cells non treated (in culture medium alone) or treated with selected H. pylori components: for 24 h with glycine acid extract – GE (10 μg/ml), urease subunite A – UreA (5 μg/ml), cytotoxin associated gene A (CagA) protein (1 μg/ml), H. pylori lipopolysaccharide (LPS Hp) or E.coli LPS –LPS Ec (25 ng/ml) or for 2 h with live H. pylori - Hp(2 × 107 colony forming units - CFU/ml), (i) intensity of fluorescence measured in a fluorescence reader at 495 nm (excitation) and 519 nm (emission), mean values ± SD. * Statistical significance for p < 0.05 assessed by non parametric U Mann-Whitney test, (ii) representative images from a confocal microscope (Leica TCS SPE) at wavelengths: FITC - 495 nm excitation and 519 nm emission, DAPI – 345 nm excitation and 455 nm emission, phalloidine – 591 nm excitation and 608 nm emission (630 × magnification). b MUC5AC and c Lewis X imaging in the gastric tissue of guinea pigs experimentally infected with H. pylori (7 and 28 days after inoculation, n = 10) or control animals (n = 5), (bi, ci) intensity of fluorescence measured using the software ImageJ version 1.48v (National Institute of Health, United States) at 495 na (excitation) and 519 nm (emission), mean values±SD. Statistical significance for p < 0.05 assessed by the non parametric Kruskal-Wallis test. * Statistically siginificant values for infected animals (7 and 28 days after inoculation) vs control animals and for infected animals 7 days post infection vs 28 days after the last inoculation with H. pylori, b ii representattive images of MUC5AC production and c ii representative images of LeX production in guinea pigs gastric tissue from fluorescence microscope (Axio Scope A1, Zeiss, Germany) at wavelengths: FITC 495 nm excitation and 519 nm emission, DAPI 345 nm excitation and 455 nm emission, (100 × magnification)

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