Novel Levamisole Derivative Induces Extrinsic Pathway of Apoptosis in Cancer Cells and Inhibits Tumor Progression in Mice
Figure 4
Expression of apoptotic proteins in CEM cells after 4a treatment.
CEM cell lysate was prepared following treatment with 4a (0, 0.5, 1 and 5 µM for 48 h). DMSO treated cells were used as control (0 µM). Western blotting studies were performed using specific primary and secondary antibodies for expression of (A) Phospho p53, p53, PUMA, phospho AKT, AKT (B) BCL2, BCL-xL, BAX and t-BID; (C) FAS, FAS-L, FADD, and SMAC/DIABLO (D) CASPASE-3, CASPASE-8 and CYTOCHROME C. α-TUBULIN was used as loading control. The quantification of the bands in each blot shown in left panel is shown as bar diagram with standard error based on two independent experiments following normalization with respective TUBULIN E. Release of CYTOCHROME C from mitochondria after treatment with 4a. Mitochondrial as well as cytosolic fractions were separated from CEM cells after 48 h of treatment with 4a (5 µM), DMSO treated cells were used as control (C), western blotting was performed using anti-CYTOCHROME C. Actin was used as loading control.