Fig. 3: DNR toxicity is increased by P2X7RA and reduced by P2X7RB expression.

a The panel represents the percentage of viable AML blasts characterized by HIGH-P2X7RA expression (AML 1 and AML 2) and HIGH-P2X7RB expression (AML 3 and AML 4) after 6 h of treatment with either vehicle (PBS, black) or DNR (200 nM, red). b, c HEK MOCK, HEK P2X7RA, and HEK P2X7RB were seeded at a concentration of 2 × 10⁵ and treated with vehicle (PBS, black) or DNR (200 nM, red) for 48 h. b The panel represents fold increase normalized on time 0 in cell numbers at 24 and 48 h after treatment. Data are presented as mean ± SEM. n = 26 microscopic fields for each group. Treatment with DNR causes a significant decrease in cell numbers for all cell types at both time points except for HEK P2X7RB at 24 h versus HEK P2X7RB at 48 h where the numbers of cells are not statistically different. These data are not depicted in the figure to increase clarity. Reported significance is relative to the comparison among different cell types all treated with DNR **P ≤ 0.01; ****P ≤ 0.0001. c Cell viability was assessed also by Alamar Blue assay, see “Materials, subjects, and methods.” 20,000 cell/100 µl were seeded in a 96-well plate and treated with vehicle (PBS, black) or DNR (200 nM, red) for 24 h. n = 18 for each group. Data are represented as mean ± SEM. *P ≤ 0.05; **P ≤ 0.01. d Extracellular ATP (pM) was measured in the culture supernatants as described in “Materials, subjects, and methods.” HEK MOCK, HEK P2X7RA, and HEK P2X7RB cells were treated for 24 h with vehicle (PBS black) or DNR (200 nM red). Data are represented as mean ± SEM. n = 6 for each group, *P ≤ 0.05; ***P ≤ 0.001,****P ≤ 0.0001.