Supplemental data

Supplemental file (.xls). Numeric dataset for all figures. Supplemental figure 1 (.pdf). Representative micrographs of rat (A) and human (B) enteric glia cultures considered as pure after immunocytochemistry (more than 90% S100β-positive cells). Cells were stained using anti S100b (green) aSMA (magenta) and antibodies. Dapi staining (blue) is shown. Scale bar, A-B, 100 mm. Supplemental figure 2 (.eps). Gating strategy for determination of live CD8 T cells. The gating strategy to first eliminate debris is shown on a representative plot SSC-A vs FSC-A. Then, single cells are identified by plotting FSC-H against FSC-A. Singlet proportion is usually in the 95-100% range. CD3 positive cells were selected by gating high B530 laser parameter. Then, positive CD8 from CD3 positive cells were selected by gating high YG780 laser parameter. Finally, live CD4 positive cells were counted by gating low V450 laser parameter. Supplemental figure 3 (.eps). Micrographs of adhesion of human CD4 and CD8 T cells derived from peripheral blood mononuclear cells on enteric glia. Enteric glia was pretreated 24 hours with IL-1b and TNFa (B-E) or not (A). D and E represent a high magnification of CD4 and CD8 cells and glial cells corresponding to the condition in which enteric glia was treated with IL-1b and TNFa and solvent. Cells were stained using anti CD4 (white) and or CD8 (magenta) and S100b (green) antibodies. In some panel, Dapi staining (blue) is shown. Scale bar, A-C, 60 mm; D-E, 12 mm.” White and yellow arrowheads point out CD4 and CD8 T cells, respectively. For more clarity, all cells are not pointed out on the figure.