Fig. 3.
Endogenous ATAD3A expression correlates with radiosensitivity in GBM cells. (A) Cell viability of wild-type T98G, wild-type U87MG, and T98G/ATAD3Akd cells by colony-forming assay after different radiation dose treatments. The wild-type T98G, U87MG, and T98G/ATAD3Akd cells were exposed to irradiation (0, 3, 6, or 12 Gy) and incubated for 10 days until colonies appeared. Viability was assessed following staining of colonies by crystal violet. Cell viability was evidently reduced in T98G-ATAD3Akd cells, suggesting that ATAD3A expression correlated with radiosensitivity. (B) Validation of silencing of ATAD3A expression (ATAD3Akd) by immunoblot. Reduced cell viability of ATAD3Akd cells after irradiation with and without TMZ (as measured by colony-formation assay). ○, T98G/EV cells; •, T98G/ATAD3Akd cells. Adding 50 μM TMZ radiosensitized T98G/ATAD3Akd (▴) but not T98G/EV (▵) cells. (C) Validation of enforced ATAD3A expression (ATAD3Aee) by immunoblotting. Radioresistance by ATAD3Aee was seen in U87MG cells after irradiation with and without TMZ. ▪, U87MG/EV cells; □, U87MG/ATAD3Aee cells. TMZ radiosensitized U87MG/EV cells (♦) but did not affect cell viability of U87MG/ATAD3Aee cells (¯). (D) The expression between ATAD3A and DDH is highly correlated, as shown by immunoblotting. GSC, glioma stem cell. (E) Knockdown of ATAD3A reduced DDH expression in T98G cells.