Fig. 1

ANRIL promoted HR repair to enhance cancer resistance in lung cancer cells. A: Representative images of the clonogenic survival assay of ANRIL NC or ANRIL-knockdown H1299 cells after 0, 2, 4, 8 Gy irradiation. B: Quantitative analysis of the clonogenic survival assay results of ANRIL-KD H1299 cells that received the indicated IR treatment. Cells transfected with shNC served as controls. Error bars represent the SEM of the mean of 3 independent experiments, two tailed Student’s t test. **P < 0.01. C: Cell apoptosis was measured with flow cytometry via the Annexin V and PI double staining method in ANRIL NC and ANRIL-KD cells at 24 h after 8 Gy irradiation. Error bars represent the SEM of the mean of 3 independent experiments, two tailed Student’s t test. **P < 0.01. D-F: Representative images from the comet assay of ANRIL-knockdown or control cells at 8 h after 8 Gy irradiation. The tail DNA percentage (E) and tail moment (F) were quantified from comet assay images in ANRIL-KD or NC cells. Error bars represent the SEM of the mean of 3 independent experiments, two tailed Student’s t test. **P < 0.01. G, H: Images and quantitative results of the γH2AX foci assay of NC and ANRIL-KD cells at the indicated time points after 8 Gy irradiation. Error bars represent the SEM of the mean of 3 independent experiments, two tailed Student’s t test. **P < 0.01. I: Representative images and of Western blotting of RPA2 phosphorylation, Rad51 phosphorylation, Chk1 phosphorylation, Chk2 phosphorylation and Kap1 phosphorylation in ANRIL-knockdown cells after irradiation. J: quantitative analysis of RPA2 phosphorylation and ATR phosphorylation in ANRIL-NC and ANRIL-KD cells. ANRIL NC cells were used as controls. The data are shown as the mean ± SEM. Significance was determined with Student’s t test. **P < 0.01, *P < 0.05. K: volumes grow curves of tumors isolated from the NC and ANRIL-KD groups with/without irradiation. Data are shown as the mean ± SD, n = 9, two-tailed Student’s t test. ***P < 0.001. L-M: Representative images of immunochemically stained Rad51 (L) and γH2AX (M) in ANRIL NC and-KD tumors at 0, 8, and 24 h after local irradiation (n = 9). N: Quantitative analysis of the percentages of RAD51-, TUNEL- and γH2AX-positive cells from IHC images from the indicated groups. Data are shown as the mean ± SD, n = 9, two-tailed Student’s t test. **P < 0.01, *P < 0.05