Volume 129, Issue 1, Pages 131-141 (July 2005) Mechanisms Underlying the Maintenance of Muscle Hypercontractility in a Model of Postinfective Gut Dysfunction  Hirotada Akiho, Yikang Deng, Patricia Blennerhassett, Hiroshi Kanbayashi, Stephen M. Collins  Gastroenterology  Volume 129, Issue 1, Pages 131-141 (July 2005) DOI: 10.1053/j.gastro.2005.03.049 Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 1 IL-4 receptorα (IL-4Rα) and 2 distinct forms of mouse TGF-β type II receptor (TGF-βR) expression in primary cultured longitudinal smooth muscle cells from control mice. This Figure represents 1 of 5 separate experiments that gave similar results. Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 2 Dose response curve of the carbachol-induced jejunal smooth muscle cell contraction from control mice (○), infected mice at day 7 PI (•), and day 35 PI (□). Values are the means ± SE of 4–6 experiments. *Significant difference of the cells from infected mice at day 7 PI compared with cells from control mice at the same concentration (P < .005). +Significant difference of the cells from infected mice at day 35 PI compared with cells from control mice at the same concentration (P < .01). CCh, carbachol. Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 3 Dose response curve of the carbachol-induced jejunal smooth muscle cell contraction from noninfected control (○), incubated with 10 ng/mL of IL-4 (•), and 10 ng/mL of IL-13 (■). Values are the means ± SE of 4–6 experiments. +Significant difference of the cells incubated with IL-4 compared with control cells at the same concentration (P < .05). *Significant difference of the cells incubated with IL-13 compared with control cells at the same concentration (P < .05). Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 4 (A) Semiquantitative TGF-β1 mRNA expression in muscle layer from noninfected control mice, infected mice at day 10, and day 28 PI. Values represent the means ± SE of TGF-β1 to G3PDH ratio from at least 3 mice. *Significant difference from correspondent noninfected controls (P < .05). (B) TGF-β1 protein quantification by ELISA in muscle layer from noninfected control mice, infected mice at day 7 and day 35 PI. The results are expressed as concentration of picograms of TGF-β1-protein per milliliter of sample solution. Values represent the means ± SE from at least 5 mice. *Significant difference from correspondent noninfected controls (P < .05). Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 5 TGF-β1 protein quantification by ELISA in primary cultured muscle cells from control, incubated with 100 ng/mL of IL-4 and 100 ng/mL of IL-13. The results are expressed as concentration of picograms of TGF-β1 protein per milliliter of sample solution. Values represent the means ± SE from at least 3 mice. *Significant difference from correspondent nonincubated controls (P < .01). Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 6 (A) COX-2 protein expression by Western blot in primary cultured muscle cells from control (lane A), incubated with 100 ng/mL IL-4 (lane B), 100 ng/mL IL-13 (lane C), or 100 ng/mL TGF-β1 for 16 hours (lane D). Each lane was loaded with 30 μg protein. These blots are representative of similar results obtained at least 3 times. (B) PGE2 protein quantification by ELISA in primary cultured muscle cells from control, incubated with 100 ng/mL IL-4, 100 ng/mL IL-13, and 100 ng/mL TGF-β1. The results are expressed as concentration of picograms of PGE2-protein per milliliter of sample solution. Values represent the means ± SE from at least 3 mice. *Significant difference from correspondent nonincubated controls (P < .05). Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 7 Dose response curve of the carbachol-induced jejunal smooth muscle cell contraction from nonincubated control (○), incubated with 0.1 nmol/L PGE2 (•), and 0.1 nmol/L PGE2 + 10−5 mol/L of celecoxib (■). Values are the means ± SE of 6 experiments. *Significant difference of the cells incubated with PGE2 compared with control cells at the same concentration (P < .05). +Significant difference of the cells incubated with PGE2 compared with cells incubated with PGE2 + celecoxib at the same concentration (P < .01). Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 8 Dose response curve of the carbachol-induced jejunal smooth muscle cell contraction from nonincubated control (○), incubated with 1 ng/mL TGF-β1 (□), and 1 ng/mL TGF-β1 + 10−5 mol/L celecoxib (■). Values are the means ± SE of 4–7 experiments. *Significant difference of the cells incubated with TGF-β1 compared with control cells at the same concentration (P < .05). +Significant difference of the cells incubated with TGF-β1 compared with cells incubated with TGF-β1 + celecoxib at the same concentration (P < .05). Gastroenterology 2005 129, 131-141DOI: (10.1053/j.gastro.2005.03.049) Copyright © 2005 American Gastroenterological Association Terms and Conditions