Extracellular DNA Containing (dG)n Motifs Penetrates into MCF7 Breast Cancer Cells, Induces the Adaptive Response, and Can Be Expressed

Figure 4

The exposure of MCF7 cells to pEGFP and pEGFP-Gn leads to an increase in ROS production. (a) FL-plate reader. The cells were analyzed in the 96-well plate format at and . The cultivation medium was replaced with 5 μm H2DCFH-DA in PBS solution, 50 ng/mL GC-DNA was immediately added, and a fluorescence intensity increase was detected at 37°C. , —sample’s signal at time and immediately after GC-DNA addition, respectively. (b) RT-qPCR. The levels of RNA NOX4 in the cells exposed to pEGFP or pEGFP-Gn for 1.5 h and 24 h. The data are normalized to the content of RNA NOX4 in the control. (c) FCA. (1) Cell plots: FL2 (NOX4-PE) versus FSC. : gated area. (2) Proportions of NOX4+ cells in gate. (3) Median signal intensity of FL2-NOX4. (d) FM-based evaluation of NOX4 (PE) in the cells treated with pEGFP-Gn or pEGFP (1.5 h). Magnification ×60. pEGFP-Gn (2): for DNA analysis of the selected area of the photo (1), the image size and brightness were increased by computer processing. .

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