Figure 2.

UNC1999 induced partially ATG7-dependent autophagy. A. LoVo, HCT-15 and DLD-1 cells were pretreated with 2 and 5 mM 3-MA for 1 h, and then exposed to 2.5 μM UNC1999 for 24 h. The protein expressions were analyzed by Western blots. B. HCT116 and DLD-1 cells were pretreated with 5 mM 3-MA for 1 h, and then exposed to 5 μM GSK343 for 24 h. The protein expressions were analyzed by Western blots. C. DLD-1 cells were transfected with ATG5 siRNA for 48 h, and then exposed to 2.5 μM UNC1999 for 24 h. The protein expressions were analyzed by Western blots. *nonspecific band. D and E. ATG7-KO and ULK1-DN DLD-1, as well as their control wildtype (WT) cells were treated with indicated doses of UNC1999 for 24 h. The protein expressions were analyzed by Western blots. F. ATG7-WT and ATG7-KO cells were treated with 5 μM UNC1999 for 24 h in the absence or presence of 20 nM bafilomycin A1 (post-treatment for 4 h). The protein expressions were analyzed by Western blots. G. ATG7-WT and ATG7-KO cells were treated with indicated doses of UNC1999 for 72 h. The cell viability was analyzed by an MTT assay. *P < 0.05 indicates significant differences between ATG7-WT and ATG7-KO cells.