Cell - 8 September 2016

Figure S4.gdT Cells Do Not Directly Modulate Pancreatitis or Transformed Epithelial Cells, Related toFigure 3
(A) Acute pancreatitis was induced using caerulein in C57BL/6-Trdctm1Malmice, which express GFP exclusively ingdT cells. Pancreata were harvested at 12h and
immunohostochemistry for GFP was performed. Arrows indicate GFP+cells.
(B) Pancreata and spleens of WT mice undergoing caerulein-induced pancreatitis were assessed by flow cytometry for the presence of CD3+TCRg/d+cells. The
percentage ofgdT cells among the intra-pancreatic or spleen T lymphocyte populations, respectively, was calculated at 48h after commencing caerulein
treatment (n = 5/group; ****p < 0.0001).
(C) WT mice were administered caerulein for up to 48h and then serially observed for a maximum additional 48h. Cohorts were sacrificed at 0 (untreated),24, 48,
72, or 96 hr from commencing caerulein treatment and the percentage of pancreas-infiltrating CD4+T cells andgdT cells among CD3+T cells was assessed by
flow cytometry (n = 3 mice/time-point).
(D–G) WT and Tcrd–/–mice were induced to develop caerulein pancreatitis for 48h. (D) Severity of pancreatitis was assessed by H&E staining, (E) CD45+pan-
leukocyte IHC, and (F) serum amylase and (G) lipase levels (n = 5/group). Pancreatitis experiments were repeated more than 5 times with similar results.
Representative H&E- and CD45-stained sections are shown.
(H–J) KPC-derived PDA cells were co-cultured with PDA-infiltrating or spleengdT cells in a 1:5 ratio for 24h. (H) Tumor cell proliferation was measured using the
XTT assay. (I) Expression of tumor suppressor or oncogenic proteins was assessed by Western blotting. (J) Expression of tumor-modulatory cytokineswas
determined in a cytometric bead array. Co-culture experiments were repeated 3 times with similar findings.