Figure 5
From: Hepatitis C virus core protein induces apoptosis-like caspase independent cell death
Influence of the HCV- core protein on caspase activation. 1 × 105 cells/ml of the UC (A, C, E) and UHCV cell lines (B) or 1 × 106 UC cells (D) were cultured for the indicated times in the presence or absence of Tet to induce specific protein expression and the broad spectrum caspase inhibitor zVAD (100 μM). The apoptotic stimuli mitomycin C (50 μg/ml), etoposide (400 ng/ml), TRAIL (40 ng/ml), and anti-CD95 antibody (100 ng/ml) were added during the last 24 h of the culture. The broad spectrum caspase inhibitor zVAD was added at day 0 if indicated. A-C: Induction of apoptosis was assessed after 48 h by flow cytometric analysis of propidium iodide staining of hypodiploid apoptotic nuclei. The mean values and standard deviation of triplicate cultures are shown (A+B). D: Cleavage of caspases-3 and -8 as well as of PARP was detected in the cell lysates by Western Blot analysis. E: Detection of the caspase activity in UC cells was performed by in vitro cleavage of the fluorogenic substrate DEVD-AMC and was measured by fluorometry at the time indicated. Apoptotic stimuli were added for 24 h. The mean values and standard deviation of triplicate cultures are given.