Changes of KEAP1/NRF2 and IKB/NF-κB Expression Levels Induced by Cell-Free DNA in Different Cell Types
(a) Intracellular location of the р65 component of NF-κB during cell incubation with GC-rich model DNA (p(rDNA)) and a plasmid vector carrying ribosomal repeats (pBR322) in a concentration of 50 ng/ml. The magnification was ×40; the exposure duration for NF-κB is indicated in the figure. (b) Fraction of cells containing NF-κB in the nucleus. The data were obtained using the Image 6 computer program for image analysis. The expression level of NF-κB was calculated in relation to the control cells cultivated without adding cfDNA fragments. The control cells were taken as a unit. . The experiment was conducted on two MSC cultures. For each culture, multiple measurements (three or more) were performed by different technicians. (c) Reads of the CyTell cell imaging system (GE Healthcare). The red sectors indicate the fraction of NF-κB-positive nuclei, while the blue sectors indicate the fractions of NF-κB-negative cell nuclei. The time of cultivation with p(rDNA) fragments and the vector (50 ng/ml) are indicated in the figure. (d) Intracellular location of NRF2 during cell incubation with oxidized cfDNA fragments (gDNAoxy) and genomic DNA (gDNA) in a concentration of 50 ng/ml. The exposure duration for NRF2 was 1 hour; the magnification was ×100.