Figure 4.

Brilliant Blue G (BBG) prevents adenosine triphosphate (ATP)‐induced cation uptake into human and murine leucocytes. (a) Human RPMI8226 and murine RAW264.7 cells, (b) human peripheral blood mononuclear cells (PBMCs) or (c) non‐obese diabetic‐severe combined immunodeficiency‐interleukin (NOD‐SCID‐IL)‐2Rγ^null (NSG) splenocytes in NaCl medium were preincubated for 15 min at 37°C in the absence or presence of (a) BBG as indicated or (b,c) 1 μM BBG. Cells were then incubated with 1 μM YO‐PRO‐1²⁺ in the absence or presence of 1 mM adenosine triphosphate (ATP) for 10 min at 37°C. The mean fluorescence intensity of YO‐PRO‐1²⁺ uptake into (a) RPMI8226 or RAW264.7 cells, (b) hCD3⁺ T cells or (b) NSG splenic CD11c⁺ dendritic cells (DCs) was then assessed by flow cytometry. (a) ATP‐induced YO‐PRO‐1²⁺ uptake is represented as a percentage of maximal ATP response in the absence of BBG. (a–c) Data represent group mean ± standard error of the mean (s.e.m.) (n = 3); *** P < 0·0001 compared to respective basal.