Extended Data Fig. 4: PARP inhibition modulates the phenotype of differentiating macrophages.
a, Schematic representation of ex vivo differentiation of CD14+ human monocytes. b, Gating strategy used in flow cytometric analysis of ex vivo differentiated human monocytes. c-e, GM-CSF plus IL-4 differentiation of ex vivo cultured macrophages treated with vehicle or Olaparib. c, There was no change in proportion of CD45+ cells, CD11b+ cells or DCs (CD11b(neg)). d, Olaparib significantly increased CD11b(neg) (dendritic cell) expression of pTBK1. e, M-CSF differentiation of ex vivo cultured macrophages treated with vehicle or Olaparib. Olaparib did not change the proportion of macrophages (CD11b+) or dendritic cells (CD11b(neg)). The frequency of cells expressing CSF-1R increased after Olaparib treatment. Data represent n = 5 human donors. Error bars represent standard error of mean (±SEM). Statistical analyses were performed using two-tailed t-test. f-g, CD14+ cells from healthy human donors were isolated and differentiated to mature myeloid cells with IL-4 and GM-CSF for 5 days at which point Olaparib was added for 4 additional days. Cells were then collected for immunophenotyping by flow cytometry f, Schematic representation of ex vivo differentiation of CD14+ human monocytes to mature macrophages. g, Olaparib did not affect the viability of mature macrophages in ex vivo cultures as shown by total viable cells. No significant changes were observed in the phenotypic markers after Olaparib was added on the differentiated macrophages. Statistical analysis was performed using unpaired one-tailed t-. Error bars represent standard error of the mean (±SEM) with n = 5 healthy human donors. Exact p values indicated in each panel for each comparison.
