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. 2007 Jan;170(1):356–365. doi: 10.2353/ajpath.2007.060736

Figure 4.

Figure 4

Effect of PKC inhibitors on MMP-9 expression and activity. A: Western blot. Western blot was performed with 100 μg of secreted proteins. MMP-9 activity determined using zymography with 25 μg of conditioned media protein. B: Western blot. Western blot analysis was performed with 400 nmol/L siRNA against PKC-α or PKC-δ alone or in combination and PMA alone (100 nmol/L). C: Gelatin zymography. Zymography was performed to test the effectiveness of PKC-α siRNA, PKC-δ siRNA alone, or in combination on MMP-9 gelatinase activity. D: Cell invasion assay. The invasion assay of control and PMA-treated HP75 cells through type IV collagen was performed to determine number of invaded cells. After 6 hours of incubation of cells with PMA, invaded cells were stained with 0.1% w/v crystal violet solution and photographed and counted. The number of invaded cells was used for subsequent comparative analyses. **P < 0.001. The effect of pharmacological inhibitors (Gö6976, BIM, and Rottlerin) of PKC and neutralizing MMP-9 antibody was determined on PMA-evoked increase in invasion. The effect of PKC-α, PKC-δ, siRNA alone, or in combination was determined on PMA-evoked increase in invasion. All treatments were performed in the presence of dimethyl sulfoxide (0.1% v/v), the vehicle in which PMA was dissolved.