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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

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

Fig. 6

A549 cells grown as 3D models confirm the irradiation-induced SASP characterized by high synthesis of DNA:RNA hybrids structures. a Representative images of SA-β-gal staining performed on FFPE tissue sections of A549 spheroids fixed after exposure to 10 or 20 Gy. For evaluation of cells positive to SA-β-Gal assay, a homemade Matlab tool was used. The appropriated mask needed to identify the cells positive to SA-β-Gal assay in the investigated area was obtained using the function imageSegmenter; (* p < 0.05; *** p < 0.0001) (b) mRNA expression of SASP biomarkers detected in A549 cells grown as 3D spheroids using GAPDH and HPRT as housekeeping genes. Data are presented as mean ± SD. c, d Confocal immunofluorescence images representative of A549 spheroids after different irradiation doses showing cytoplasmic (green) and nuclear (white) localization of DNA:RNA hybrids (scale bar 10 μm). The micrograph shows the co-localization analysis performed through confocal high-resolution analysis. Bar graphs, mean ± error bars of fluorescent signals in the different compartments are also reported (see Methods section) (* p < 0.05)

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