1. Volume 134, Issue 2, Pages 459-469 (February 2008)
Protein Kinase C θ Plays a Fundamental Role in Different Types of Chronic Colitis 
Kiyotaka Nagahama, Atsuhiro Ogawa, Katsunori Shirane, Yasuyo Shimomura, Ken Sugimoto, Atsushi Mizoguchi 
Gastroenterology 
Volume 134, Issue 2, Pages (February 2008)
DOI: /j.gastro
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2. Figure 1
Localization of PKCθ to IS in CD4+ T cells of inflamed colon. Colonic CD4+ T cells from CD45RB model (upper panel) or WT mice (lower panel) were stained with anti-PKCθ (green) and choleratoxin (red) and subjected to confocal microscopic analysis.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

3. Figure 2
Contribution of PKCθ in CD4+ T cells for the induction of a Th1-mediated colitis. (A and B) Representative gross appearance (A) and histologic findings (B) of the colon of the RAG1KO mice with transfer of naïve CD4+ T cells from PKCKO or WT mice are shown. (C) Colitis scores of recipient RAG1KO mice at 8 weeks after the cell transfer with T cells from PKCθKO or WT mice are summarized, *P < (D) The expressions of IL-2/IL-6 and IFN-γ/IL-17A in the gated CD3+CD4+ colonic T cells that were stimulated with anti-CD3/CD28 mAbs are shown. These data are representative of 3 individual experiments. (E) The absolute numbers of colonic WT (black bars) or PKCθKO (gray bars) CD4+ T cells that produce IL-2, IL-6, IFN-γ, and IL-17A in the recipient colon are summarized.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

4. Figure 3
Requirement of PKCθ for the development of Th2-mediated colitis. (A and B) Representative gross appearances (A) and histologic findings (B) of the colon of TCRαKO mice (A, upper panels) and αPKCDKO mice (A, bottom panels) are shown. (C) Disease scores of TCRαKO vs αPKCDKO mice at 6 months of age are summarized. There is a statistical significant difference (P < .0001) in the disease score between TCRαKO and αPKCDKO mice. (D) Total RNA of 3 × 105 purified colonic CD4+ T cells from TCRα (black bars) or αPKCDKO (gray bars) mice were subjected to real-time PCR analysis for measurement of GATA3, IL-4, IL-13, and IL-2.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

5. Figure 4
Requirement of PKCθ-signaling pathway for the expansion of colonic CD4+ T cells under intestinal inflammatory conditions. (A–D) The proportions (A and C) and absolute numbers (B and D) of CD4+ CD3+ T cells in the colonic LP of recipient RAG1 KO mice (CD45RB model), after being reconstituted with WT (A, left panel) or PKCθ-deficient (A, right panel) CD4+ T cells, and from TCRαKO (C, left panel) and αPKCDKO mice (C, right panel) are shown (A and C). (E) The proliferative responses of colonic CD4+ T cells from recipient RAG1 KO mice, after being reconstituted with WT or PKCθ-deficient CD4+ T cells, and from TCRαKO and αPKCDKO mice are shown. The representative data of 3 individual experiments are shown. (F) Annexin-V staining of colonic CD4+ T cells from recipient RAG1KO mice (CD45RB model), after being reconstituted with WT or PKCθ-deficient CD4+ T cells, and from TCRαKO and αPKCDKO mice is shown. The data are representative of 2 individual experiments.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

6. Figure 5
Involvement of PKCθ in the development of colitis that is induced by absence of IL-2. (A and B) Representative gross appearance (A) and histologic findings (B) of the colon of βIL2DKO mice reconstituted with CD4+ T cells from WT mice (top) or PKCθKO mice (bottom) are shown. (C) Colitis scores of βIL2DKO mice without or with transfer of CD4+ T cells from PKCKO or WT mice are summarized. *P <
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

7. Figure 6
Involvement of PKCθ in Th2 cytokine expression and in the proliferation of Th1 and Th17 subsets. (A) Expressions of IL-17A, RORγt, IFN-γ, T-bet, IL-2, and IL-6 by the purified CD4+ T cells from the colon of WT mice (open bars, n = 3) and βIL2DKO mice that are being reconstituted with CD4+ T cells from WT (black bars, n = 7) or PKCθKO (gray bars, n = 7) mice are shown. *P < .05, **P < (B) FACS analysis shows the expression of CD4 vs IL-17A or IFN-γ expressions on colonic lamina propria cells from recipient βIL-2DKO mice reconstituted with CD4+ T cells from WT (left panels) or PKCθKO mice (right panels) after stimulation with anti-CD3/CD28. (C–E) βIL2DKO mice, after being reconstituted with WT or PKCθKO CD4+ T cells (C and D), TCRαKO mice, and αPKCDKO mice (E) were injected IP with 0.1 mg of BrdU on 48 and 24 hours prior to death. Colonic LP cells were then stimulated with plate-coated anti-CD3 mAb/soluble anti-CD28 mAb and then stained for detection of IL-17 and IFN-γ (C and D) or IL-4 (E) in CD4+ BrdU+ vs CD4+BrdU− T cells. The percentages of IL-17+ /IL-17− and IFN-γ+ /IFN-γ− in the gated BrdU+ CD4+ T cells are summarized in D.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

8. Figure 7
Contribution of PKCθ in the proliferation but not AICD of CD4+ T cells in the absence of IL-2. (A) The proportions and absolute numbers of CD3+ CD4+ T cells in the spleen and colon of βIL2DKO mice reconstituted with WT or PKCθ-deficient T cells are shown. (B and C) Expression of CD62L vs CD44 on gated CD3+ CD4+ T cells from the spleen and colon (B) and expression of CD69 on colonic CD4 + T cells (C) from recipient βIL2DKO mice reconstituted with WT or PKCθ-deficient T cells are shown. As control, expression of CD62L vs CD44 on the gated CD3+ CD4+ T cells from the spleen of WT mice is shown in the left panel. (D) Apoptotic responses (as judged by Annexin-V staining) in the splenic and colonic CD4+ T cells from recipient βIL2 DKO mice reconstituted with WT or PKCθ-deficient T cells are shown. The data (n = 8/each group) are summarized in the right panel. (E) Western blot analysis shows the comparable expression levels of Bcl-2, Bax, and β-actin in the purified colonic CD4+ T cells from βIL2DKO mice reconstituted with WT or PKCθ-deficient CD4+ T cells. (F) Proliferative responses (as judged by BrdU incorporation) of CD4+ T cells from the spleen and colon from recipient βIL2DKO mice reconstituted with WT (n = 3) or PKCθ-deficient (n = 3) T cells are shown.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions