Figure 4.

Caspase-3 activation by H₂O₂ and STS treatment in uninfected and HIV persistently-infected lymphoid cell lines. A) H9, H9/HTLVIII_B, Jurkat and J1.1 cells were exposed to H₂O₂ and STS. After 24 h, cells were washed and lysed with RIPA buffer. Equal amounts of protein (30 μg/sample) were separated on a 10% SDS-PAGE and blotted onto nitrocellulose membranes. Blots were probed with anti-procaspase-3 for 1 h and revealed with a peroxidase-conjugated anti-IgG antibody and ECL (enhanced chemoluminiscence) Equal loading was checked by analyzing β-actin expression (data not shown). B) H9 and H9/HTLVIII_B cells were exposed to 10μM H₂O₂, 0.1 μM STS or complete medium for 24 h and collected to evaluate active caspase-3 by PE-conjugated monoclonal anti-active caspase-3 antibody by flow cytometry. C) Jurkat and J1.1 cells were exposed to 10 μM H₂O₂, 0.1 μM STS, 40 ng/ml CH11 or complete medium for 24 h and collected to evaluate active caspase-3 by PE-conjugated monoclonal anti-active caspase-3 antibody by flow cytometry as described in Materials and Methods