Fig. 4. Kaempferol and cisplatin co-treatment induced oxidative stress-mediated inflammatory response in human colon cancer HCT-15 and HCT-116 cells. Colon cancer cells treated with kaempferol, and cisplatin combination promotes changes in reactive oxygen species (ROS) production which is observed by alterations in fluorescence observed via different staining. (A, B) Representative fluorescent microscopy images of HCT-15 and HCT-116 colon cancer cells stained with H2DCFDA. (C, D) Visualization of mitochondrial ROS was carried out with MitoSOX^TM Red staining in HCT-15 and HCT-116 cells. Scale bar of images 100 μm. The graphs represent the relative fluorescence intensity of the cells treated with kaempferol and cisplatin co-treatment as compared to control. (E) Kaempferol and cisplatin co-treatment further increased the level of nitric oxide production in cells. (F) HCT-15 and (G) HCT-116; kaempferol and cisplatin combine treated cells altered the redox signaling. Quantification of H2DCFDA, MitoSOX^TM Red fluorescence intensity and relative protein expression intensity were analyzed with ImageJ software. The data are represented as the mean ± standard deviation. *P < 0.05, **P < 0.01, and ***P < 0.001 for comparison between control and treated cells, whereas ^#P < 0.05, ^##P < 0.01, and for comparison between kaempferol alone and combination and ^$P < 0.05, ^$$P < 0.01 for comparison between cisplatin only and combine treatment. ns, not significant.

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