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STEM_599_sm_suppinfofigS1.tif3003KSupplementary Figure 1. Ets2 deficiency alters the differentiated state of colon crypts. (A) The thickness of the mucosa from sections of V-Cre, R26R mice of varying Ets2 genotype and age stained for beta-galactosidase were measure. Mucosal thickness was measured at 6-8 different lenghts along the colon. Bars represent the average of the measurements from all mice or the corresponding genotype and age and error bars represent the standard deviation. Solid black bars represent Ets2flox/flox mice while solid gray bars represent Ets2flox/+ mice. For crypt depth at 15 days two Ets2flox/flox, and Ets2flox/+ were inspected. For 30 days n= 4 and 4. For 60 days n=3 and 2. For 90 days n= 3 and 1. (B) goblet cell abundance at 30 days of age. The number of goblet cells and the total number of cells were counted in crypts from 5 different fields taken at 200×. Bar graphs of the number of globlet cells divided by total nuclei multiplied by 100, is presented as the percentage of goblet cells. P value is from Mann-Whitney test. (C) total cell number /crypt. Cell numbers were determined from 42 crypts from 4 Ets2flox/flox mice and 24 crypts from 3 Ets2flox/+ mice. Uniformly blue crypts were scored in the Ets2flox/flox mice while a blue and white crypts were scored in the Ets2flox/+ mice. (D) Crypt density is increased in mice with Ets2 deficient crypts. Stretches of 10 or more adjacent, well organized crypts were identified in beta-galactosidase stained sections. The number of crypts was divided by the length of the measured area. At least 3 different area were measured per mouse. Stretches exclusively containing uniformly blue crypts were scored in the Ets2flox/flox mice while streches containing blue and white crypts were scored in the Ets2flox/+ mice.
STEM_599_sm_suppinfofigS2.tif3099KSupplementary Figure 2. Tumor from WT and AA mice do not differ in proliferation, apoptosis or inflammatory cell involvement. (A) Histological comparison of late wt and AA colon tumors. Upper panel: Hematoxylin and eosin stained sections of colon tumors from WT and AA mice, second panel: anti-PCNA staining, third panel: TUNEL staining and bottom panel: anti-F4/80 staining. Three upper panels: scale bar represents 200 μm, bottom panel: scale bar represents 50 μm. (B) Quantitation of anti-PCNA staining. Tumor areas were encircled using the ellipse tool in scanscope, and the number of PCNA positive pixels were calculated within the area. Bar graphs represent the number of positive pixels per area in mm2. N represents the number of tumors examined for each genotype. (C) Quantitation of TUNEL staining. Bar graphs represent the number of positive pixels per area in mm2. Ctrl represents a negative control section (no primary antibody). (D) Quantitation of anti-F4/80staining Bar graphs represent the number of positive pixels per area in mm2. Ctrl represents a negative control section (no primary antibody).
STEM_599_sm_suppinfotable1.doc65KSupplementary Table 1: Analysis of tumor size and grade.

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