Fig. 3

WZ35 exhibits significant antitumor activity. A CCK-8 assays were utilized to measure the suppression rate of curcumin and WZ35 (with the concentration of 0, 5, 10, 20 to 40 μg/mL) treated HCCLM3, HepG2, and Huh7 cells. B Colony formation assays of HCCLM3, HepG2, and Huh7 cells were conducted by plating 5000 indicated cells per well in 6-well plates to measure the effect of curcumin and WZ35 on clonogenicity with representative images and quantification of colony numbers being shown. C Representative fluorescence microscopy displaying nuclear transformations in HCCLM3, HepG2, and Huh7 cells following 18 h treatment of WZ35 (10 μg/mL) or treatment of curcumin (10 μg/mL) by means of DAPI staining. Results are presented as the mean ± standard error from independent experiments performed in triplicate. *P < 0.05, **P < 0.01, student’s t test. D The levels of various apoptosis associated proteins, including Bax, Bcl-2, and Cleaved caspase-3 were detected by western blotting following the 18 h treatment of WZ35, with GAPDH serving as an internal control. E Quantitation of the murine body weight and tumor volumes over time in four indicated tumor xenograft groups, as measured by utilizing electronic balance and caliper. The results are presented as the mean ± standard error from independent experiments in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001, student’s t test. F Subcutaneous tumor xenograft models of WZ35-treated HCCLM3 were used. Mice were put to death by cervical dislocation and the tumors were harvested after 17 days. Optical image of each tumor and the weight curve was photographed and quantified, respectively. (n = 6 mice per group). G) The representative images of Hematoxylin and eosin staining of indicated treated tumors have been shown. Scale bars, 200 μm for images