Fig. 3

CRT is exposed at the cell surface and ATP is released. a Apotome immunofluorescence detection of CRT using CRT polyclonal antibodies (red) at hour 24 of treatment with βGBP (2 nM) and confirmation of CRT cell surface location by co-staining with the fluorescent lectin wheat germ agglutinin (green). Scale bars: 10 μm. Images are representative of at least three independent experiments. b Detection of CRT at the cell surface by flow cytometry using CRT polyclonal antibody at hour 24 of treatment with βGBP (2 nM). Right peaks represent percent expression of CRT in treated cells (black line) and in controls (grey line). Left peaks (grey infill) represent non-specific antibody binding: isotype controls. Data are representative of at least three independent experiments. c Time course of CRT expression on SW620 and SW480 βGBP and mock treated cells. Propidium iodide (PI) exclusion was used to stain dead cells. Values are means of three independent experiments  ± SD. d ATP release at 24 and 48 h by 2 × 10⁵ SW620 and SW480 cells as percentage of total cellular ATP. Black histograms mock treated cells, white histograms βGBP-treated cells (2 nM). ATP release assessed with ATPlite assay. Values are means of triplicate experiments  ± SEM