Fig. 7: Response of primary human monocytes and NK cells to supernatants from TNBC cells treated with the autophagy inhibitor SAR405 or radiation. | Cell Death & Disease

Fig. 7: Response of primary human monocytes and NK cells to supernatants from TNBC cells treated with the autophagy inhibitor SAR405 or radiation.

From: Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition

Fig. 7

PBMCs from three different donors were incubated for 20 h in the presence of the culture medium (control) or cell-free supernatants from A MDA-MB-468, B MDA-MB-436 and C MDA-MB-231 cells. A MDA-MB-468 cells were treated with 3 μM SAR405 for 96 h, 3×8 Gy X-rays or a combination thereof. Supernatants were harvested 96 h after the last irradiation. When combined with radiation, SAR405 was added 96 h before harvesting. B MDA-MB-436 cells were irradiated with 8 Gy X-rays or C-ions and harvested after 96 h. C MDA-MB-231 cells were treated with 3 μM SAR405 for 96 h. Human monocytes (left) and NK cells (right) were identified according to lineage markers and their activation was assessed based on the analysis of activation markers CD80, CD86, HLA-DR, CD25 (monocytes), CD69, CD25, CD38 (NK cells) or activation-induced molecules CD39 and PD-1 with immunoregulatory functions (NK cells) by spectral flow cytometry. Representative histograms of one donor (top) and percentages of marker-positive cells ± standard deviation of three PBMC donors (bottom) are shown. In histograms, the gating for the positive cells is based on the lineage stain (depicted in gray) and is indicated by a vertical line. Experiments in A and C were performed at the same time, thus they share the medium control. Statistical significance was assessed using one-way ANOVA with Tukey’s post-hoc test (*≤ 0.05; **≤ 0.01; ***≤ 0.005, ****≤0.0001).

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