Fig. 4: TOPK decreases ULK1 activity in glioma cells.

a TOPK-overexpression-U118MG cells were harvested at 80% confluence. ULK1, ATG13, p-ATG13, Beclin-1, p-Beclin-1, HA, and Tubulin were analyzed using western blots. b TOPK-silencing-Hs683 (left) and TOPK-silencing-H4 (right) cells were harvested at 80% confluence. ULK1, ATG13, p-ATG13, Beclin-1, p-Beclin-1, TOPK, and Tubulin were analyzed using western blots. c Hs683 (left) and H4 (right) cells were treated with different concentrations of OTS964 for 48 h, and the samples were resolved by SDS–PAGE and analyzed by western blots using the indicated antibodies. d TOPK-silencing-Hs683 (left) and TOPK-silencing-H4 (right) cells were treated with EGF (20 ng/ml) for 15 min followed by the addition of CHX (100 μg/ml) for 6 and 8 h before extraction. ULK1 and Tubulin were analyzed using western blots. e Hs683 (left) and -H4 (right) cells were treated with or without OTS964 (1 μM) for 48 h, then cells were exposed to EGF (20 ng/ml) for 15 min followed by the addition of CHX (100 μg/ml) for 6 and 8 h before extraction. ULK1 and Tubulin were analyzed using western blots. f H4 cells were treated with or without OTS964 (1 μM) for 48 h, and then cells were exposed to CHX (100 μg/ml) for 6 and 8 h before extraction. TOPK and Tubulin were analyzed using western blots. Data represent the results of triplicate experiments