Fig. 2

Effects of MFX and CFX on biochemical events associated with apoptosis. a As described in material and method, caspase-8, 9, 3 activities were measured in MIA PaCa-2 (i), and Panc-1 cells (ii), in presence and absence of MFX/CFX for 48 h. The enzyme activity was measured by extent of cleavage of the caspase substrates Ac-IETD-pNA, Ac-LEHD-pNA and Ac-DEVD-pNA respectively. Bar graph represents the mean ± SEM of the fold increase in enzyme activity versus untreated control of three independent experiments performed in duplicates. Here vertical axis represents fold change in caspase activity. *p < 0.015, #p < 0.05 b Western blot analysis of Bid activation and PARP cleavage in MIA PaCa-2 (i), and Panc-1 cells (ii), treated with MFX/CFX in a dose dependent manner for 48 h. GAPDH was used as loading control. Data are representative of typical experiment repeated three times with similar results. Bar Graph represents the mean ± SEM. here vertical axis represents fold change and horizontal axis represents concentration in μg/ml. *p < 0.01 versus control. c DNA was isolated from MFX/CFX treated MIA PaCa-2 (i), and Panc-1 cells (ii) for 48 h, as described in material and method section, and was resolved onto 1.8 % agarose gel to detect DNA fragmentation, the characteristic feature of cells undergoing apoptosis. Pictures are representative of three independent experiments. (1) represents standard DNA marker, (2) DNA from untreated cells, (3) cells treated with 100 μg/ml of MFX, (4) cells treated with 200 μg/ml of MFX, (5) cells treated with 400 μg/ml of MFX, (6) cells treated with 100 μg/ml of CFX, (7) cells treated with 200 μg/ml of CFX, (8) cells treated with 400 μg/ml of CFX