BBT activates JNK-, p38-,
and caspase-dependent apoptosis. (A)
Cell cycle histograms of Hela cells after treatment with various concentrations
of BBT. Analysis of apoptosis using the annexin V and PI method. (B)
HeLa cells were incubated in the absence (control) or presence of
BBT (12.5 μM). Then, cell death was analyzed using annexin V
and PI through flow cytometry. (C) Flow-cytometric analysis of mitochondrial
membrane potential (ΔΨm) with the help of JC-1 staining
in treated and untreated samples.(D) Immunoblotting analysis of cell
lysates collected from HeLa cells after treatment of 12.5 μM
BBT. Stress response pathway (p-JNK1/2, p-c-jun, and p-p38) and apoptotic
factors (cleaved caspase 9 and 3 and PARP) were seen to be upregulated
on treatment with BBT. α-Tubulin was used as a loading control.
All experiments have been performed thrice (n = 3).
PI: propidium iodide, PARP: poly(ADP-ribose) polymerase, and JNK:
Janus kinase.