Fig. 5
Biological functions of PINT87aa. a Left, cell cycle was determined in PINT-87aa or circPINTexon2 stably overexpressed 456 and 4121 BTICs. Right, cell cycle analysis of PINT87aa K.O SW1783 and Hs683 cells. b MTT proliferation assay was examined in PINT87aa-GFP or circPINTexon2 stably overexpressed 456 and 4121 cells or PINT87aa K.O SW1783 and Hs683 cells. c Edu proliferation assay was examined in PINT87aa-GFP or circPINTexon2 stably overexpressed 456 and 4121 cells, PINT87aa K.O SW1783 and Hs683 cells and their control cells. d Soft-agar and plate colony assay was performed in PINT87aa K.O SW1783 and Hs683 cells and their control cells. Scale bar, 100 μM. e In vitro extreme limiting dilution assays (ELDAs) were performed to evaluate BTICs self-renewal capacity. Scale bar, 100 μM. Spheres were counted at 14 days. Cell density per well ranged from 1, 10, 25, 50, 100, 250, 500 to 1000. Each condition was tested in 10 independent wells. Neurosphere-forming capability was determined using the ELDA web-based tool (456, P < 0.001; 4121, P < 0.001, by ELDA analysis). f PINT-87aa or circPINTexon2 stably overexpressed 456 and 4121 BTICs and their control cells were subjected to 6 Gy radiation. DNA damage was determined by flow cytometry and γ-H2AX expression. a–d, f Data are presented as mean ± s.e.m. from three independent experiments. *P < 0.05; ns, P > 0.05, determined by two-tailed Student’s t-tests
