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. Author manuscript; available in PMC: 2015 Sep 10.
Published in final edited form as: Mol Carcinog. 2013 Oct 22;53(0 1):E169–E180. doi: 10.1002/mc.22088

Figure 2.

Figure 2

Reg4 expression in murine GI tract and various CRC cell lines parallels with that of β-Catenin and TCF-4 transcription factor. (A) Immunohistochemical staining of tissues from different regions of murine (BALB/c) GI tract exhibited a parallel between Reg4 and β-Catenin expression. Comparatively higher levels of Reg4 in cecum and colon corresponded to higher levels of β-Catenin in similar regions, whereas the lower level of Reg4 in duodenum paralleled with lower β-Catenin expression. (B) Following Western blot analyses similar parallels between Reg4 and β-Catenin expression were also observed in different human CRC cells. Compared to HCT116 cells, SW480 and HT29 cells showed higher levels of Reg4 and β-Catenin protein and were associated with increased protein expression of TCF-4 transcription factor. (C) Compared to non-transfected 293 cells, 293-R4 cells (293 cells stably transfected with Reg4 clones) showed increased protein levels of β-Catenin and TCF-4 transcription factor. (D) Compared to HCT116 cells, higher levels of Reg4 in SW480 and HT29 cells were also associated with increased TCF-4 activity assessed by TOP/FOP ratio using luciferase reporter assay system. HCT116 cells showing comparatively lower expression of Reg4 exhibited lower TOP/FOP ratio. TOP/FOP ratio in 293 human embryonic kidney cells with non-detectable level of Reg4 protein served as negative control.