Figure 2.

ConA triggers autophagic but not apoptotic cell death in human hepatoma cells. (a) HuH-7 and Hep G2 cells were treated with ConA (20 μg/ml) or Puro (1 μg/ml) in the presence or absence of either Baf A1 (1 nM) or Z-VAD (20 μM) for 24 h. After treatment, cytotoxicity was analyzed by LDH release assay. (b) HuH-7-shLuc, HuH-7-shLC3, HuH-7 wild-type (WT) and HuH-7 ATG9 knockout (KO) cells were treated with different doses of ConA as indicated for 24 h. After treatment, cytotoxicity was analyzed using an LDH release assay (c) HuH-7 cells were treated with ConA (20 μg/ml) or Puro (1 μg/ml) in the presence or absence of Z-VAD (20 μM) for 24 h. Cell lysates were subjected to detect pro-caspase 3 and cleaved-caspase 3. (d) HuH-7 cells were treated with ConA (20 μg/ml) or Puro (1 μg/ml) in the presence or absence of Z-VAD (20 μM) for 24 h. Sub-G1 phase was performed by fixed PI staining and analyzed using flow cytometry. Quantification of sub-G1 phase was analyzed using the WinMDI 2.9 software (the upper left). All data are presented as the mean±S.D. from at least triplicate independent experiments; *P<0.05, **P<0.01, ***P<0.001, ns indicates no significance