Skip to main content
Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: TP53 drives abscopal effect by secretion of senescence-associated molecular signals in non-small cell lung cancer

Fig. 7

EVs secreted by high-dose irradiated A549 cells induce in vitro abscopal effects. a EVs secreted by irradiated A549 stimulate the synthesis of DNA:RNA hybrid structures. Immunofluorescence staining with S9.6 antibody. The images are representative of unIR A549 cells exposed to EVs isolated from culture medium of A549 cells non irradiated or irradiated at different doses. Cell images were captured by Nikon Eclipse Ti2 confocal microscope with 60x plan apochromat oil immersion objective lens. Data are presented as grand median ± MAD. b EVs secreted by irradiated A549 induce senescent phenotype. (I) Representative images of SA-β-gal staining of UnIR A549 cells after exposure to EVs isolated from culture medium of IR A549 cells. II. Colony-forming assay (CFA). The cells were exposed to EVs isolated from UnIR or IR A549 cells for 14 days. 0.05% crystal violet solution was used to visualize the generated colonies with more than 50 cells, which were quantified under inverted microscope (Olympus IX51 microscope, Olympus Corporation, Tokyo, Japan) by two independent observers. III. Expression levels of senescence markers. p21, IL6 mRNA levels were measured by Real-Time PCR and normalized to GAPDH and HPRT-1. Data are the mean of two independent experiments. (C) EVs secreted by irradiated A549 induce M1 polarization in a murine macrophage cell line. Total RNA from RAW 264.7 M0 cells (see Methods section) exposed to EVs secreted by unirradiated, 10 Gy or 20 Gy-irradiated A549 cells were analyzed by RT-PCR for the expression of representative murine M2 genes (Arg1, Egr2) and M1/pro-inflammatory cytokines (IL-6, IL-1β). Expression data are given as fold increase over the mRNA level expressed by RAW 264.7 M0 exposed to UnIR EVs. Data are represented as mean ± SD of triplicate values of two independent experiments (*p < 0.05, **p < 0 .01 ***p < 0.001)

Back to article page