Figure 8

Effect of SNP and PTIO on calreticulin exposure in HT29 iNOS^- cells. HT29 iNOS^- cells and HT29-dx cells were incubated for 3 h in the absence (CTRL) or in the presence of PTIO (100 μmol/L), SNP (100 μmol/L) or both (SNP+PTIO) for 3 h, and then the surface CRT was investigated by the following procedures. (A, B, C) FACS analysis of surface CRT. The negative controls, with non-immune isotypic antibodies, are shown by green outline in panel A and B. The figures shown here are representative of three similar experiments, done in triplicate. In panel C the percentage of cells positive for surface CRT is represented as mean ± SE in all experiments. Versus CTRL: * p < 0.05; versus SNP: ° p < 0.05. (D, E) Western blot detection of surface CRT and total CRT in HT29 iNOS^- cells and HT29-dx cells after a biotinylation assay (see Methods for details). Total CRT was detected in an aliquot of cells incubated under the same experimental conditions of the biotinylation assay, subjected to ultracentifugation and Western blot analysis. The results shown here are representative of two similar experiments, with superimposable results.