Samples were placed into an automated tissue processor (TBS, model ATP1) and allowed to process overnight. 3 day reversal to water. F) Distal colon wall thickness and G) mucosa layer/crypt length after exposure to the various DSS protocols. Data are reported as Mouse monoclonal to ERBB3 group means SEM. Asterisks correspond to p-values as follow: * p 0.05, **p 0.01 & ***p 0.001. Sample size for each group is usually indicated within its bar around the RK-287107 bar graph.(TIF) pone.0220156.s001.tif (14M) GUID:?DA06D188-BF39-459F-9E8A-A9BA5BD39A72 S2 Fig: Rating scales utilized for the subjective assessment of distal colon inflammation and goblet cell loss. Description of common RK-287107 pathology corresponding to each score on subjective rating scales for any) inflammation and B) goblet cell loss with representative images for each score.(TIF) pone.0220156.s002.tif (15M) GUID:?C028D852-1E85-47A1-AC17-CDE293DC76D9 S3 Fig: Differential response in the open field by mice exposed to 4% DSS for 4 days or 7 days with or without reversal to water. A) Total number of rears after 60 moments in the open field test revealed significant suppression in mice exposed to DSS for 4 and 7 days and then reverted to water for 3 extra days, but not in mice exposed to DSS for 4 days. B) Assessment of total ambulation revealed that, of the groups tested, only the 4 day DSS plus 3 day reversal to water group exhibited decreased movement. C) Normalizing rears by dividing rear counts by overall RK-287107 movement revealed that only the 7 day DSS plus 3 day reversal group exhibited decreased normalized rears. D) Computation of the average time spent per rear revealed that both 4 and 7 days of DSS exposure plus reversal to water, but not 4 day RK-287107 DSS exposure, caused significantly shorter rearing bouts than controls. Data are reported as group means SEM. Asterisks correspond to p-values as follow: *p 0.05, **p 0.01, ***p 0.001. Sample size for each group is usually indicated within its bar on the bar graph.(TIF) pone.0220156.s003.tif (2.9M) GUID:?F9D64109-EB77-4C90-B288-E4CD864EC4A1 S4 Fig: Colon inflammation was present by the 4th day of DSS exposure and remained present even in mice that were reverted to regular drinking water for 3 days. A) Distal colon tissue expression levels of TNF were significantly elevated after 4 days of exposure to DSS. B) IL-1 colon expression was greater in animals exposed RK-287107 to DSS for longer periods (7 days), and remained significantly elevated even after 3 days reversal to water. C) MPO and D) NPY expression levels were generally higher in mice exposed to DSS than controls and tended to be higher with longer exposure time. Data are reported as group means SEM. Asterisks correspond to p-values as follow: *p 0.05, **p 0.01, ***p 0.001. Sample size for each group is usually indicated within its bar on the bar graph.(TIF) pone.0220156.s004.tif (2.8M) GUID:?D3148ECF-9ACB-4F05-975B-31974D1D2A78 S5 Fig: Combined systemic treatment with the Y1 (BIBP 3226) and the Y2 (BIIE 0246) receptor inhibitors did not further improve DSS-induced pathology relative to either antagonist alone. All mice were exposed to a 4% DSS answer for 7 days and treated with either BIBP 3226, BIIE 0246 or a cocktail made up of both antagonists injected once daily. A) Percent body weight switch in DSS uncovered mice exhibited that Y2 receptor inhibition significantly slowed down excess weight loss progression. B) H&E stained distal colon sections demonstrating DSS-induced histopathology and immune cell infiltration. C) Colon length and D) muscle mass wall thickness were not altered by combined treatment with the antagonists. E) Rears normalized to locomotion were comparable between groups. F) Average rear duration was significantly decreased by Y1 receptor inhibition, increased by Y2 receptor inhibition and unchanged in the combined inhibitor treatment group. G) Distal colon tissue expression levels of TNF were comparable between groups whereas H) IL-1 levels were significantly suppressed by Y1 receptor antagonism and to a comparable degree, in the group receiving the combined treatment. Similarly, L) DSS-stimulated.