Fig. 3: TEMs/TAMs suppress olaparib-induced DNA damage in BRCA1-deficient breast tumor cells.

a Diagram of workflow for (b)–(f). Conditioned media from control macrophages (Mφ) or tumor-educated macrophages (TEMs) were added to tumor cell cultures, followed by olaparib treatment. b BP cells were stained using DAPI and an anti-dsDNA antibody after two days of olaparib treatment. The intensity of dsDNA fragments in the cytosol was quantified. Scale bar, 50 μm (control, n = 21 fields; olaparib, n = 26 fields; control macrophages, n = 10 fields; control macrophages + olaparib, n = 10 fields; TEMs, n = 14 fields; TEMs + olaparib, n = 10 fields examined over two independent experiments). c BP cells were stained with anti-H2AX phospho-Ser139 antibody and analyzed by flow cytometry. MFI, median florescence intensity (n = 3). d BP cells were analyzed for apoptosis (Annexin V⁺ 7-AAD⁻) (n = 4). e MDA-MB-436 cells were stained with anti-H2AX phospho-Ser139 antibody and analyzed by flow cytometry (n = 4). f MDA-MB-436 cells were analyzed for apoptosis (Annexin V⁺ 7-AAD⁻) (n = 4). g Apoptotic analyses of BP cells co-cultured with or without TAMs sorted from BP tumors, followed by three days of olaparib treatment (n = 4). Flow cytometry plot axes are displayed in logarithmic scale. h Schematic representation of the experiments for (i) and (j). i and j Analysis of TAMs (i) and γ-H2AX in CD45-negative cells (j) in BP tumors after two IP injections of anti-CSF1R antibody and 7 days of treatment with olaparib (OL) as monotherapy or in combination. Control, n = 4; OL, n = 6; anti-CSF1R, n = 4; OL + anti-CSF1R, n = 6. Each dot represents data from a single tumor. Data are presented as mean ± SEM (b)–(g) or median with quartiles (violin plots, i and j). One-way ANOVA. ns, not significant. Source data are provided as a Source Data file.