Fig 1: The Wnt target gene Troy can be detected in gastric cancer cell lines and overexpression of variant 2 suppresses the tumor phenotype of MKN45 cellsA. Expression profile of Troy and its variant 2 in different cell lines compared with non-neoplastic human stomach tissue [n=3 for non-neoplastic tissue (NT) and tumor (TU), n=2 for cell lines]. Expression was normalized to SDHA in all cell culture experiments. B. Expression of Troy in MKN45 cells is not significantly changed after 5azaC treatment for 72 h (n=3). C. qRT- PCR analysis of Troy and LGR5 expression after stimulation of MKN45 with 10 nM Wnt3a (n=2) alone or in combination with 10 nM RSPO1 and/or RSPO2 (n=3-4). D. - E. MTT based proliferation analysis of stable transfected MKN45 (D) and MKN74 (E) overexpressing Troy variant 2 compared to control transfected cells (n=3). F. Colony formation assay evaluates the ability for clonal expansion of Troy variant 2 transfected MKN45 (n=8). All data are represented at mean +/- SD with *p=0.051; **p=0.108; ***p=0.031).
Fig 2: Expression of LGR5 and TROY in mice during weeks post treatment. The arrows demonstrate the inverse trend
Fig 3: Immunohistochemical detection of Troy+ cells in the gastric corpus mucosaA-D. Whole section overview of the stomach epithelium. As expected, nuclei were immunonegative (original magnification A: 60x, inserts B-D: 250x). E-I. Identification of Troy+ cells (brown DAB staining) by double staining with lineage markers (VectorRed): gastric H+/K+-ATPase (E), pepsinogen C (F), chromogranin A (G), Muc6 (H) or Muc5 (I); original magnifications 400x. K. Troy+ myocytes allowed for internal staining control.
Fig 4: Representative slides of mouse gastric tissue. (A) H&E staining (scale bars: 200 µM), (B) immunodetection of LGR5 (scale bars: 50 µM), and (C) TROY (scale bars: 50 µM) expression at different time points post treatment
Fig 5: Expression of LGR5 and TROY mean composite scores (CS) of (A) tumor and non-tumoral adjacent tissues and (B) relative expression (CStumor/CSadjacent) of LGR5 and TROY. The arrows in panel A depict the inverse trend
Supplier Page from Abcam for Anti-TROY antibody [EPR3213(2)]