FIGURE 6:

LanCL2 depletion sensitizes cells to apoptosis through down-regulating Akt phosphorylation. (A) HepG2 cells were infected with lentiviruses expressing LanCL2 shRNA (“L-1” and “L-2”) or control (“C”), selected with puromycin, and then treated with 10 ng/ml TNFα and 10 μg/ml CHX for 3.5 h or 5 μM adriamycin for 17 h; this was followed by Western analysis. (B) Cells treated with TNFα and CHX as in A were subjected to TUNEL assays. Data shown are the average of three independent experiments, and ∼30,000 cells were examined for TUNEL signals for each data point. (C) Lentivirus-infected and puromycin-selected cells were transfected with HA-myr-Akt or a control plasmid; this was followed by TNFα/CHX treatment as in A. The cells were fixed and immunostained for cleaved PARP and HA. The percentage of transfected cells positive for cleaved PARP was calculated. Data shown are the average of three independent experiments. A paired t test was performed to compare each data point with the control. *, p < 0.05.