Fig. 5

Ebselen suppresses autophagy flux via inhibition of ATG4B. A, B HeLa cells were treated with Torin 1 (2 μM) or CQ (40 μM) and detected by western blot. C, D SW620 cells were transduced with sgCon and sgATG4B lentiviruses, respectively. Then cells were treated with CQ (40 μM) and detected by western blot. E, F HCT116 were treated with Ebselen (5 and 10 μM) or CQ (40 μM) and subjected to western blot. G, H SW620 were treated with Ebselen (10 μM) or CQ (40 μM) and western blotting was carried out to detect the protein levels. I, J Immunofluorescence analysis of endogenous LC3B in HCT116 treated with Ebselen (10 μM) or CQ (40 μM). Dots numbers for LC3B were derived from counting at least 20 cells and 5 fields from three independent replicated experiments. Bar = 10 μm. K HCT116 cells were treated with 2 μM Torin 1 in the presence or absence of 10 μM Ebselen for 12 h, then the level ratio of long-lived proteins was measured by flow cytometry