1. Volume 125, Issue 1, Pages 148-161 (July 2003)
Colonic epithelial functional phenotype varies with type and phase of experimental colitis 
Emiko Mizoguchi, Ramnik J Xavier, Hans-christian Reinecker, Hirofumi Uchino, Atul K Bhan, Daniel K Podolsky, Atsushi Mizoguchi 
Gastroenterology 
Volume 125, Issue 1, Pages (July 2003)
DOI: /S (03)

2. Figure 1
Histological analysis of DSS-induced colitis and TCRα KO mice and the purification method of colonic epithelial cells. Histological analysis of distal part of colons of (A) WT, DSS-induced colitis on (B) day 4, (C) day 8, and (D) day 12 and (E) TCRα KO mice is shown (objective, 20×). (F) Isolated colonic crypts from WT mice by 30 mmol/L EDTA perfusion through the left ventricle were stained with H&E (objective, 20×). After collagenase type IV digestion, contaminating hematopoietic cells were sequentially removed by negative selection with monoclonal Ab-coated magnetic beads. The purified cells were stained with anti-TROMA-1 Ab, which specifically reacts with epithelial cells (objective, 40×).
Gastroenterology  , DOI: ( /S (03) )

3. Figure 2
Down-regulated gene expression of detoxification and biotransformation related molecules of CECs in murine colitis models. (A) Transcription of SBP, CAR-IV, PST-1 and Mdr1a, carbonyl reductase (CBR), and β-actin mRNA of purified CECs from C57BL/6 WT, DSS administrated C57BL/6 on day 4 (4d), day 8 (8d), and day 12 (12d), and TCRα KO mice with colitis were examined by RT-PCR. (B) Transcription of SBP, CAR4, PST-1, Mdr1a, and β-actin messenger RNA of purified CECs from C57BL/6 WT, IL-10 KO mice, and CD45RB model was examined by RT-PCR. (C) The expression ratio of each messenger RNA level evaluated by densitometry analysis (Quantity one; Bio Rad) is compared with WT mice. The ratio of WT mice is represented as 1.0. The data represent mean values of 6 mice in each group ± standard error of the mean. ∗P < 0.01; ∗∗P < versus WT mice. m represents 100 base pair (bp) molecular weight marker.
Gastroenterology  , DOI: ( /S (03) )

4. Figure 3
Tissue localization and role of CAR under inflammatory conditions. (A) Western blot analysis of purified mouse CAR-IV fusion protein tagged with Xpress was detected by rabbit serum (lanes 1 and 2), HRP conjugated anti-Xpress antibody (lanes 3 and 4), and rabbit antibovine CAR polyclonal antibody (lanes 5 and 6). (B, C) Immunohistochemical analysis with anti-CAR antibody in DSS-induced colitis on (B) day 4 and (C) TCRα KO mice with severe colitis is shown (objective, 20×). (D-G) Mdr1a staining of colon in DSS-induced colitis on (D) day 0 and (E) day 4 and in (F) TCRα KO mice with (F) mild and (G) severe colitis is shown (objective, 20×).
Gastroenterology  , DOI: ( /S (03) )

5. Figure 4
Effect of inhibiting detoxification-related molecules in DSS-induced colitis. (A) Body weight change (percentage compared with the initial body weight before starting DSS treatment) of DSS colitis in C57BL/6 WT mice receiving intrarectal administration of PBS as control (○), valproic acid (■), and niflumic acid (□). The administration of inhibitors was performed on days −1, 0, 1, 2, 4, and 6. The values represent the mean ± standard error of the mean of 10 mice in each group. (B) Clinical score of WT mice treated with PBS (○), valproic acid (■), and niflumic acid (□). The values represent the average of 10 mice in each group. (C) Representative gross appearance of colon of DSS-colitis mice (day 12) treated with PBS (left) or valproic acid (right). (D, E) Representative photo of BrdU-incorporated CEC in (D) PBS-treated mice and (E) valproic acid-treated mice on day 12 after DSS induction (objective, 10×). (F) Number of BrdU-incorporated CECs/crypt unit in valproic acid- ( ) or PBS- (■) treated mice after DSS induction. The values represent the average of 6 mice in each group. (G) Body weight change (percentage compared with the initial body weight before DSS) in C57BL/6 WT mice receiving intrarectal administration of PBS as control (open circle), valproic acid, (■) and niflumic acid (□). The administration was performed on days 5, 6, 7, 8, 9, and 10 after initiated DSS colitis. (H) Clinical score of WT mice treated with PBS (○) and valproic acid (■). ∗P < 01; ∗∗P < 0.01; ∗∗∗P < versus PBS-treated mice.
Gastroenterology  , DOI: ( /S (03) )

6. Figure 4
Effect of inhibiting detoxification-related molecules in DSS-induced colitis. (A) Body weight change (percentage compared with the initial body weight before starting DSS treatment) of DSS colitis in C57BL/6 WT mice receiving intrarectal administration of PBS as control (○), valproic acid (■), and niflumic acid (□). The administration of inhibitors was performed on days −1, 0, 1, 2, 4, and 6. The values represent the mean ± standard error of the mean of 10 mice in each group. (B) Clinical score of WT mice treated with PBS (○), valproic acid (■), and niflumic acid (□). The values represent the average of 10 mice in each group. (C) Representative gross appearance of colon of DSS-colitis mice (day 12) treated with PBS (left) or valproic acid (right). (D, E) Representative photo of BrdU-incorporated CEC in (D) PBS-treated mice and (E) valproic acid-treated mice on day 12 after DSS induction (objective, 10×). (F) Number of BrdU-incorporated CECs/crypt unit in valproic acid- ( ) or PBS- (■) treated mice after DSS induction. The values represent the average of 6 mice in each group. (G) Body weight change (percentage compared with the initial body weight before DSS) in C57BL/6 WT mice receiving intrarectal administration of PBS as control (open circle), valproic acid, (■) and niflumic acid (□). The administration was performed on days 5, 6, 7, 8, 9, and 10 after initiated DSS colitis. (H) Clinical score of WT mice treated with PBS (○) and valproic acid (■). ∗P < 01; ∗∗P < 0.01; ∗∗∗P < versus PBS-treated mice.
Gastroenterology  , DOI: ( /S (03) )

7. Gastroenterology 2003 125, 148-161DOI: (10.1016/S0016-5085(03)00665-6)

8. Figure 5
Up-regulated expression of genes encoding anti-inflammatory peptides in both acute and chronic colitis models. (A) Transcription of MRP 14, MRP8, SLPI, SOCS3, IDO, WDNM1, and β-actin messenger RNA of purified CECs from C57BL/6 WT, DSS administrated C57BL/6 on day 4 (4d), day 8 (8d), and day 12 (12d), and TCRαKO mice with colitis was examined by RT-PCR. (B) Transcription of MRP14, MRP8, SLPI, SOCS3, IDO, WDNM1, and β-actin messenger RNA of purified CECs from C57BL/6 WT, IL-10 KO mice, CD45RB model, and TCRα KO mice was examined by RT-PCR. (C) Transcription of SLPI and β-actin messenger RNA of purified CECs from TCRα KO mice and IL-10-deficient TCRαdouble KO (IL-10 × αDKO) was examined by RT-PCR. (D) The expression ratio of each messenger RNA level evaluated by densitometry analysis is compared with TCRα KO mice. The ratio of TCRα KO mice is represented as 1.0. The data represent mean values of 6 mice in each group ± standard error of the mean. ∗P < 0.01; ∗∗P < versus WT mice. m represents 100-bp molecular weight marker.
Gastroenterology  , DOI: ( /S (03) )

9. Figure 6
Protein expression of anti-inflammatory molecules of CECs in murine colitis models. (A) Western blot analysis of purified mouse SLPI fusion protein tagged with 6× His was detected by goat serum (lane 1), rabbit serum (lane 2), HRP conjugated anti-6× His antibody (lane 3, 4), and goat antihuman SLPI antibody (lanes 5 and 6). (B) Colony formation of BL21 LysS E. coli strain carrying the gene encoding IPTG-inducible SLPI was spread on the plate coated without (right) or with (left) IPTG. (C, D) Immunohistochemical analysis using anti-SLPI antibodies in DSS-induced colitis on (C) day 4 and (D) TCRα KO mice with severe colitis is shown (objective, 20×). (E, F) Immunohistochemical analysis with anti-SOCS3 antibodies in DSS-induced colitis on (E) day 4 and (F) TCRα KO mice with severe colitis is shown (objective, 20×).
Gastroenterology  , DOI: ( /S (03) )

10. Figure 7
Up-regulated gene expression of regeneration-related molecules of CECs in murine colitis models. (A) Transcription of Reg-IIIβ, -IIIγ, SLFN 2, and β-actin messenger RNA of purified CECs from C57BL/6 WT, DSS administrated C57BL/6 on day 4 (4d), day 8 (8d), and day 12 (12d), and TCRαKO mice with colitis was examined by RT-PCR. (B) Transcription of RegIIIγ, RegIIIβ, SLFN, and β-actin messenger RNA of purified CECs from C57BL/6 WT, IL-10 KO mice, CD45RB model, and TCRαKO mice was examined by RT-PCR. (C) The expression ratio of each messenger RNA level evaluated by densitometry analysis is compared with TCRα KO mice. The ratio of TCRα KO mice is represented as 1.0. The data represent mean values of 5–6 mice in each group ± standard error of the mean. ∗P < 0.01; ∗∗P < versus WT mice. m represents 100-bp molecular weight marker.
Gastroenterology  , DOI: ( /S (03) )

11. Figure 8
Protein expression of regeneration-related molecules of CECs in murine colitis models. (A) Western blot analysis of purified mouse Reg IIIγ fusion protein tagged with Xpress was detected by rabbit Ig (lanes 1 and 2), HRP conjugated anti-Xpress antibody (lanes 3 and 4) and rabbit antimouse Reg IIIγ antibody (lanes 5 and 6). (B-E) Immunohistochemical analysis with anti-Reg IIIγ antibody in DSS-induced colitis on (B) day 0, (C) day 4, (D) day 12, and (E) TCRα KO mice with severe colitis is shown (objective, 20×). (F-I) MHC class II (IAb) staining of colon in DSS-induced colitis on (F) day 4, (G) day 8, and (H) day 12, and in (I) TCRα KO mice with severe colitis is shown (objective, 20×).
Gastroenterology  , DOI: ( /S (03) )