Fig. 3. Complexes RAS-1H and RAS-1T induce early time-point ROS and activate the cellular antioxidant defence mechanism. (a) Detection of ROS with carboxy-H2DCFDA (20 μM) after treatment with RAS-1H and RAS-1T for 3 h, 6 h and 9 h using a microplate assay. Mean ± s.e.m. (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; Student's t test). (b) Detection of ROS with a fluorescence microscope after treatment for 6 h. (c) Western blot analysis of Nrf-2, a central protein in cellular antioxidant defence and (d) expression levels of the Nrf-2 target gene in AGS cells after treatment with RAS-1H, RAS-1T and cisplatin at LD and HD for 6 h and 24 h. Homogeneous protein loading determined with reference to actin and gene expression normalized against tbp levels.
