Figure 5

Detection of surface calreticulin in HT29, HT29-dx and HT29 iNOS^- cells. Cells were grown in absence (CTRL) or presence of 5 μmol/L doxorubicin (doxo) for 24 h and subjected to the following investigations. (A, B) FACS analysis of surface CRT. Negative controls, with non-immune isotypic antibodies, are shown in A by green outline. The figures shown here are representative of three similar experiments, performed in triplicate. In panel B the percentage of cells positive for surface calreticulin is represented as mean ± SE in all experiments. Versus HT29 CTRL: * p < 0.005. (C) Western blot detection of surface CRT and total CRT. The measurement of CRT associated with the plasma membrane was performed in a biotinylation assay as described in the Methods section. Under the same experimental conditions, an aliquot of cells was lysed and ultracentrifuged to detect the total CRT amount. The results shown here are representative of two similar experiments.