Inhibition of constitutive NF-κB activity by GTX and PGA1 induces QMIC apoptosis.

Human blood T cells (A), B cells (B), granulocytes (C), and monocytes (D) were isolated from buffy coats by density centrifugation and negative magnetic selection and cultured for 90 minutes in the presence or absence of 1.5 μM mGTX, 1.5 μM GTX, or 48 μM PGA1, except for monocytes, which were treated with 5 μM mGTX, 5 μM GTX, or 96 μM PGA1. Nuclear extracts were then prepared and analyzed for NF-κB–binding activity by EMSAs. The arrows indicate specific NF-κB complexes. EMSAs are representative of at least 3 comparable assays. Untreated and treated cells were also cultured for 6 hours and 24 hours (mononuclear cells) or 2 hours, 4 hours, and 6 hours (granulocytes) before conducting apoptosis assays using a dual-color annexin-V–FITC/PI staining and flow cytometry analyses. Data are presented as means ± standard deviations (n = 6).