Zymography Analysis

The results from the zymography test, shown in figure 1, confirm the results in the hypotheses. In figure 1, there are two horizontal bands labeled MMP-9 and MMP-2. In addition there are twelve lanes, with the first lane representing the marker and lanes two through 12 representing the different groups in the lab. Groups 2, 4, 6, 8, 10, and 12 completed the test with PMA treated HL-60 cells and groups 3, 5, 7, 9, and 11 completed the test with DMSO treated cells. Zymography testing shows the enzymatic activity by showing varying colored bands. During the test, the MMPs eat away the gelatin just as they would eat away the extracellular matrix if they were in a living organism and as they eat away the gelatin the color begins to fade away and
The DMSO treated cells still show some MMP-9 activity because some of these cells differentiate into monocytes, which produce MMP-9, even though most differentiate into granulocytes, which do not produce MMP-9. Monocytes are present in DMSO treated cells because DMSO is made with PMA, meaning there are trace amounts of PMA contained in DMSO. One other limitation can be seen in the differing thickness of the MMP-2 bands. This means that some of the cell samples contained more enzymatic activities, such as group 12 MMP-2 results being much thicker than the group 8 MMP-2 results. Tis could have been caused by one group loading a larger sample in than another, but this cannot be confirmed because the amount of protein cannot be seen in these results and the only thing that can be seen is the amount of enzymatic activity. Even with these limitations, the effects of differentiation on the protein can be seen because of the clear difference in MMP-9 enzymatic activity being much higher in cells treated with PMA then in cells treated with