1. Volume 133, Issue 3, Pages 918-925 (September 2007)
Enteric Glial-Derived S100B Protein Stimulates Nitric Oxide Production in Celiac Disease 
Giuseppe Esposito, Carla Cirillo, Giovanni Sarnelli, Daniele De Filippis, Francesco Paolo D’Armiento, Alba Rocco, Gerardo Nardone, Raffaella Petruzzelli, Michela Grosso, Paola Izzo, Teresa Iuvone, Rosario Cuomo 
Gastroenterology 
Volume 133, Issue 3, Pages (September 2007)
DOI: /j.gastro
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2. Figure 1
S100B immunohistochemistry in duodenal mucosa. (A) S100B immunoreactivity was mainly detectable within the submucosa of control subjects. In GFD-CD and CD patients (B and C, respectively), a stronger S100B immunopositivity was present in the submucosa, along with a S100B immunoreactivity in the mucosa (original magnification, ×100). In controls, GFD-CD, and CD-patients (Ai, Bi, and Ci, respectively), the pattern of S100B-positive enteric glia is represented at high magnification (×400).
Gastroenterology  , DOI: ( /j.gastro )
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3. Figure 2
(A) S100B mRNA expression by quantitative real-time PCR. S100B mRNA was increased in patients with CD, respect to GFD-CD patients and controls. The fold increases were relative to the control group after normalization to β-actin mRNA. Each bar shows the mean ± SD of 3 experiments, **P < .01. (B) Western blot analysis showing the expression of S100B protein in controls, CD, and GFD-CD patients. The S100B protein expression in tissue homogenates is shown in the upper panel, whereas, the densitometric analysis of corresponding bands is represented in the graph. Upper panel is representative of n = 3 experiments. Each bar in the graph shows the mean ± SD of 3 experiments. **P < .01 vs GFD-CD patients and controls. (C) Increase of S100B release by duodenal biopies from CD vs GFD-CD patients and controls. Each bar shows the mean ± SD of 3 experiments. **P < .01 vs controls and GFD-CD patients.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

4. Figure 3
(A) Western blot analysis showing the expression of iNOS protein in controls, CD, and GFD-CD patients. iNOS protein expression in tissue homogenates is shown in the upper panel, whereas the densitometric analysis of corresponding bands is represented in the graph. Upper panel is representative of n = 3 experiments. Each bar in the graph shows the mean ± SD of 3 experiments. **P < .01 vs controls and GFD-CD patients. (B) NO levels in controls, CD, and GFD-CD cultured biopsy specimens. NO production was assessed measuring the accumulation of nitrite in the culture medium at 24 hours. Each bar shows the mean ± SD of 6 experiments. **P < .01 vs controls and GFD-CD patients.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

5. Figure 4
(A) Western blot analysis showing the effects of exogenous S100B protein (0.005–5 μmol/L) at 24 hours in cultured duodenal biopsy specimens derived from controls. iNOS protein expression in tissue homogenates is shown in the upper panel, whereas the densitometric analysis of corresponding bands is represented in the graph. Upper panel is representative of n = 3 experiments. Each bar in the graph shows the mean ± SD of 3 experiments. *P < .05, **P < .01 vs untreated. (B) Effect of exogenous S100B protein (0.005–5 μmol/L) on NO production in cultured duodenal biopsy specimens derived from controls. NO production was determined by measuring the accumulation of nitrite in the culture medium at 24 hours. Each bar shows the mean ± SD of 6 experiments. *P < .05, **P < .01 vs untreated.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

6. Figure 5
(A) Effect of 6 hours incubation of exogenous S100B protein (0.005–5 μmol/L) on malonyldialdehyde levels, as marker of lipid peroxidation, in cultured duodenal biopsy specimens derived from controls. Each bar shows the mean ± SD of 6 experiments. °°P < .01 vs untreated. (B) Western blot analysis showing the effect exogenous S100B protein (0.005–5 μmol/L) on p-p38 MAPK protein expression at 6 hours in cultured duodenal biopsy specimens derived from controls. p-p38 MAPK protein expression in tissue homogenates is shown in the upper panel, whereas the densitometric analysis of corresponding bands is represented in the graph. Upper panel is representative of n = 3 experiments. Each bar in the graph shows the mean ± SD of 3 experiments. *P < .05, **P < .01 vs untreated.
Gastroenterology  , DOI: ( /j.gastro )
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7. Figure 6
Inhibitory effect of p38 MAPK inhibitor SB (0.03–3 μmol/L) (A) and NF-κB inhibitor TLCK (0.01–1 μmol/L) (B) on the production of NO induced by exogenous S100B protein (5 μmol/L) in cultured duodenal biopsy specimens derived from controls. NO production was determined by measuring the accumulation of nitrite in the culture medium at 24 hours. Each bar shows the mean ± SD of 6 experiments. **P < .01 vs untreated; °°P < .01 vs S100B-treated biopsy specimens.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

8. Figure 7
(A) S100B mRNA expression by quantitative real-time PCR. S100B mRNA was increased at 24 hours in GFD-CD specimens challenged with pt-gliadin (1 mg/mL) but not in controls. S100B mRNA levels were quantified relative to β-actin levels. No significant differences in expression of the housekeeping gene (β-actin) were found between treatment groups. Each bar shows the mean ± SD of 3 experiments. **P < .01 vs controls and GFD-CD patients. (B) Western blot analysis showing the effect of pt-gliadin (1 mg/mL) on S100B protein expression at 24 hours in cultured duodenal biopsy specimens derived from controls and GFD-CD patients. The S100B protein expression in tissue homogenates is shown in the upper panel, whereas the densitometric analysis of corresponding bands is represented in the graph. Upper panel is representative of n = 3 experiments. Each bar in the graph shows the mean ± SD of 3 experiments. **P < .01 vs untreated. (C) Increased S100B release at 24 hours by duodenal biopsy specimens from controls and GFD-CD patients cultured with pt-gliadin. No effect was observed in control biopsy specimens. Each bar shows the mean ± SD of 3 experiments. **P < .01 vs untreated.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

9. Figure 8
(A) Western blot analysis showing the effect of pt-gliadin (1 mg/mL), in the presence or absence of S100B antibody (1:100,000–1:1000, vol/vol dilution), on iNOS protein expression at 24 hours in cultured duodenal biopsy specimens derived from GFD-CD patients. iNOS protein expression in tissue homogenates is shown in the upper panel, whereas the densitometric analysis of corresponding bands is represented in the graph. Upper panel is representative of n = 3 experiments. Each bar in the graph shows the mean ± SD of 3 experiments. **P < .01 vs untreated; °°P < .01 vs untreated. (B) Effect of pt-gliadin (1 mg/mL), in the presence or absence of S100B antibody (1:100,000–1:1000, vol/vol dilution) and glial fibrillary acidic protein antibody (1:1000, vol/vol dilution), on NO production at 24 hours in cultured duodenal biopsy specimens derived from GFD-CD patients. NO production was determined by measuring the accumulation of nitrite in the culture medium. Each bar shows the mean ± SD of 6 experiments. **P <. 01 vs untreated; °°P < .01 vs untreated.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions