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. 2016 Jun 14;7(30):47232–47241. doi: 10.18632/oncotarget.10030

Figure 5. Zingerone inhibits the activities of MMP-2 and MMP-9.

Figure 5

(A) CM was collected from Renca cells that were treated with zingerone for 24 h and hypoxic conditions for 2 h. CM was subjected to gelatin-based electrophoresis and the gel was stained with Coomassie brilliant blue. (B) The activities of MMP-2 and MMP-9 were measured by quantification of bands with Image J software. **p < 0.01 and ***p < 0.001 compared to the normoxic control; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared to the hypoxic control. (C) Renca cells were co-transfected with pGL2-MMP-2 promoter and pRSV-β-galatosidase vectors. After transfection, the cells were incubated in the presence of zingerone for 24 h under hypoxic conditions. Relative luciferase activity was calculated by normalization of transfection efficiency according to β-galactosidase activity. ***p < 0.001 compared to the hypoxic control. (D) Renca cells were exposed to zingerone for 24 h and hypoxic condition for 2 h before harvesting of whole cell extracts, and Western blot analysis was performed using indicated antibodies. p-JNK (1:1000), p-NK-κB (1:1000), p-STAT3 (1:1000), p-ERK (1:1000). (E) Relative phosphorylation of JNK level was expressed as the means ± S.D. of three independent experiments. The expression under hypoxic control was set to 100%. *p < 0.05, and **p < 0.01 compared to the hypoxic control. (F) Renca cells were exposed to zingerone for 24 h and hypoxic condition for 2 h before harvesting of whole cell extracts, and Western blot analysis was performed using p-c-Jun antibody (1:1000). (G) Relative phosphorylation of c-Jun level was expressed as the means ± S.D. of three independent experiments. The expression under normoxic control was set to 100%. *p < 0.05, and ***p < 0.001 compared to the hypoxic control.