Fig. 5

DNA hypomethylation of the SMAD3 promoter region in CAFs leads to increased radioresistance of NSCLC cells. A The expression of SMAD3 in NSCLC cell lines and human bronchial epithelial cells 16HBE measured by RT-qPCR. B The expression of SMAD3 in NAFs and CAFs measured by RT-qPCR. C Western blot of SMAD3 protein in the CAF-CM and NAF-CM. D The DNA methylation level of the SMAD3 promoter region in NAFs and CAFs measured by MSP-PCR. E The expression of SMAD3 in H460 cells treated with oe-SMAD3 or CAF-CM and in H1229 cells treated with sh-SMAD3 measured by RT-qPCR. F Viability of H460 and H1229 cells treated with oe-SMAD3 or sh-SMAD3, respectively, following exposure to different doses of X-ray radiation. G Proliferation of H460 and H1229 cells treated with oe-SMAD3 or sh-SMAD3, respectively, following exposure to 6 Gy X-ray radiation. H Invasion of H460 and H1229 cells treated with oe-SMAD3 or sh-SMAD3, respectively, following exposure to 6 Gy X-ray radiation. I γH2ax fluorescence in H460 cells treated with oe-SMAD3 or CAF-CM and in H1229 cells treated with sh-SMAD3 following exposure to 6 Gy X-ray radiation. J Apoptosis of H460 cells treated with oe-SMAD3 or CAF-CM and of H1229 cells treated with sh-SMAD3 following exposure to 6 Gy X-ray radiation determined by flow cytometry. K Cell cycle distribution of H460 cells treated with oe-SMAD3 or CAF-CM and of H1229 cells treated with sh-SMAD3 following exposure to 6 Gy X-ray radiation determined by flow cytometry. *p < 0.05 vs. 16HBE, NAF, or oe-NC groups. ^#p < 0.05 vs. NAF-CM group. ^&p < 0.05 vs. sh-NC group. Cell experiments were repeated three times