Aberrant activation of nuclear factor of activated T cell 2 in lamina propria mononuclear cells in ulcerative colitis

Figure 1 Two-dimensional gel electrophoresis identifying increased amounts of NFAT2 in UC colonic mucosa tissue. Tissue protein lysate was prepared from normal (A) and UC affected colon tissues (B) and separated using two-dimensional gel electrophoresis (2-DE). Spot identification and matching across the gels and determination of the relative amount for each corresponding protein spot were conducted using the software, Progenesis workstation (non-linear). Proteins were identified using mass spectrometry as previously described[30]. The representative results of 2-DE are shown. The inserts are the close views along with the 3-dimensional pictures of NFAT2 and the reference protein spot on the 2-DE. Arrows indicate the reference protein spots across gels. The grey lines outline the NFAT2 spots.

Figure 2 Immunohistochemical analysis of the expression of NFAT2 in colon mucosa tissues. A-C are derived from a case of normal control, D-F from a case of UC, and G-I from a case of CD. A, D ,G are the results of H&E stain in 100 × magnification. B, E, H are the results of immunohistochemistry for NFAT2 counter-staining with hematoxylin for nuclei, 100 × magnification. C, F, I are the close view of B, E, H respectively. Of note, NFAT2 was exclusively located in cytoplasm of LMPCs of normal colon mucosa as well as LMPCs of CD affected colonic mucosa, whereas NFAT2 was primarily located in the nuclei of LMPCs of the UC affected colonic mucosa.

Figure 3 Confocal microscopy demonstrating the subcellular distribution of NFAT2 in LMPCs of colon mucosa. A-C are derived from normal colon mucosa, while D-F from UC affected mucosa, G-I from CD affected mucosa. A, D, G represent the detection for NFAT2; B, E, H: The nuclei using DAPI; C, F ,I : the merged images for A, B, D, E, G and H, respectively. Of interest, co-localization of NFAT2 with nuclei is noted in the colonic mucosa of UC (F), but not in CD (I), though there is a heavy infiltration of LMPCs in the CD affected colonic mucosa.

Figure 4 Nucleo-translocation of NFAT2 in UC, CD, and non-specific colitis. Nucleo-translocation of NFAT2 was assayed using immuno-histochemical staining and the tissue sections were obtained from a total of 107 cases of UC, 15 cases of CD and 3 cases of non-specific colitis (NC). Overall P value < 0.001 (determined via Kruskal-Wallis statistic).