Fig. 7

BLf induces caspases-3, −8 and −9 cleavage in breast cancer cell lines. Western blot analysis of cleaved caspases-8 and −9 in MDA-MB-231 (a) and MCF-7 (b) cells following treatment with 40 nM Apo-bLf and Fe-bLf for 24 h. Western blot analysis of pro and cleaved caspase-3 in MDA-MB-231 (c) and MCF-7 (d) cells following treatment with 40 nM Apo-bLf and Fe-bLf for 24 h. Lysates were collected and 100 μg loaded and run on standard SDS-PAGE followed by transfer to PVDF membranes. Membranes were then blocked with 1% skim milk followed by incubation with anti-cleaved caspases-3, −8 and −9 primary antibodies (Cell Signalling) and anti-mouse secondary antibody (Sigma-Aldrich). Membranes were viewed via XRS camera. Band densities determined by ImageJ software and compared with untreated (0 nM). Separate, identical gels were run for GAPDH (Santa Cruz) which were used to normalize band densities. Band density analysis was performed using ImageJ software. Fold change was calculated relative to untreated cells. Relative fold change values per band are indicated below blots as well as plotted on graphs