Figure 11
Effects of BzATP on apoptosis in cultured mouse keratinocytes. In all experiments levels of apoptosis were normalized to an arbitrary value of 2 in control cells. AU – arbitrary units. A. BzATP dose-response effect in cultured mouse normal (wild-type, C57Bl) keratinocytes (means ± SD, n = 3). Cells were treated with one of the indicated concentrations of BzATP for 8 hours. B. Cultured mouse normal keratinocytes (wild-type, C57Bl) were pre-treated with 100 μM anti-sense P2X7 oligonucleotides (ASO) or random-control P2X7 oligonucleotides (RCO) for 14 hours followed by 8 hours treatment with 100 μM BzATP. Control – cells treated with the vehicle of the ASO. Values are means (± SD) of 3 experiments for each condition. Insert in B is Western immunoblot with anti-P2X7 antibody of lysates of cells treated with ASO or RCO (n = 2). C. BzATP time-response effect in cultured mouse normal keratinocytes (wild-type, C57Bl; filled triangles), or in keratinocytes obtained from P2X7-deficient (P2X7-/-Pfizer [empty circles] or P2X7-/-GSK [filled circles]) mice (both in the C57Bl background). Values are means (± SD) of 3 experiments for each condition. Changes in apoptosis in A and B were determined in terms of solubilized DNA; changes in apoptosis in C were determined using cell-death detection ELISA.