Effect of BT on pro-apoptotic (pP38) and pro-survival (pAkt, NF-κB) signalling molecules. (A) Cells were treated with 50 μM or 100 μM BT for 24 hrs, proteins extracted, subjected to electrophoresis and detected by western blotting using primary antibodies specifically recognizing the phosphorylated active forms of these proteins. As an internal standard for equal loading, blots were probed with an anti β-actin antibody. (B) Effect of p38 inhibitor SB203580 and pAkt inhibitor LY294002 on BT treated cells. To assess role of p38 and pAkt activation in BT induced cytotoxicity, cells were treated with 100 μM BT in presence of 10 μM of SB203580 or 10 μM LY294002 (non-toxic conc.’s) for 48 hrs and cell viability determined by presto blue cell viability reagent. The results represent % viability when compared with untreated cells. Data was expressed as mean ± SD of triplicate experiments. C. Effect of BT treatment on NF-kB regulated cell survival genes such as pIkBα, xIAP, pbcl-2, bcl-xL, as analyzed by western blotting of cellular lysates.