Data Collection:
An MTS Assay was conducted on the microtiter plate with the Colo cells as well as the MCF7 cells. In each plate, the first row (row A) was filled up only with medium and contained no cells, providing a base value of absorption when conducting the MTS Assay. This first row contained dilutions of the drug and medium only. It is important to note that there was some precipitate present at the higher concentrations of IP6 (0.037mM and 3mM), but this precipitate was minimal and was ignored.

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Since this experiment was done in duplicate, the results from two columns were averaged to find the average absorption for a certain concentration. This average was calculated starting from row B to row H, and then the average of the corresponding columns in row A were subtracted from this average. This gives the absorption of the samples minus the background absorbance.
For example, in the Colo data, in row B for the concentration of 0mM of IP6, the average of 0.654 and 0.711 are taken. The average of the corresponding cells in row A (0.233 and 0.213) are also averaged and then subtracted from the average of the row B results.

These values are then plotted onto the graphs Figure 3a(i) and Figure 3b(i).

Data Analysis:

Refer to Figure 3a(i).
In Figure 3a(i), there results showed that there was first a slight increase in cell absorbance of the MTS Assay, before the drop in absorbance at the higher concentrations. These results somewhat verify that the IP6 is limiting the cell proliferation. Furthermore, there is also a decrease of the MTS Assay as the concentration of the PG490 was increased, showing that there was less cell proliferation. This verifies that an increased concentration of PG490 will limit the proliferation of Colo cells. There seems to be a somewhat steady decrease between the concentrations 0.4ng/mL and 50ng/mL.

However, the two drugs are not proven to be synergetic, just additive. They are not necessarily helping each other. To further investigate this, another set of graphs showing the cell viability with each drug must be plotted.
Refer to Figure 3a(ii).
From the results, another graph was compiled where the viability of the cells was compared against the concentration of IP6, without PG490. The 100% viability mark is where there was no drug used to treat the cells. The point where 50% viability is achieved, the IC50, will be noted. This graph is based on the absorbance of the LNCaP
cell samples. In this graph, with the cells with IP6 alone, a smooth curve line of best fit is drawn to determine the IC50.

The following graph was compiled where the viability of the cells was compared against the concentration of PG490, without IP6. The 100% viability mark is where there was no drug affecting the results. The point where 50% viability is achieved, the IC50, will be noted.
Refer to Figure 3a(iv).
In this graph, a smooth curve line of best fit can also be drawn and the IC50 can be obtained. The IC50 is determined to be 1.8ng/mL of PG490.

With these IC50 values, another graph is compiled where the viability of the cells treated with the IC50 of PG490 when plotted against increasing concentrations of IP6. Since the IC50 of the PG490 was 9.6ng/mL the closest concentration to this that was used was 10.0ng/mL.
Refer to Figure 3a(iii)
In the graph, the IC50 cannot be determined once again and is still greater than 3.0mM like before. However, whether or not the IC50 was affected is impossible to tell. The curve of the graph changed, usually when this happens there is synergy, but it is inconclusive.

With these IC50 values, another graph is compiled where the viability of the cells treated with the IC50 of IP6 when plotted against increasing concentrations of PG490. Since the IC50 of the IP6 was greater than 3mM, the closest concentration to this that was used was 3mM. The viability of the cells with 3mM of IP6 influencing their growth is plotted against increasing concentrations of PG490.
Refer to Figure 3a(v).
In this graph, the IC50, is determined to be 6.5ng/mL, which is similar to the IC50 of the cells with increasing PG490 concentrations without the influence of IP6. As a result, with no change in IC50, the PG490 does not effect the IP6 synergistically.

Refer to Figure 3b(i).
In Figure 3b(i), the results show that the absorbance of the MTS Assay decreased at the higher concentrations of IP6, ignoring the last column. These results verify that IP6 is limiting the cell proliferation. Furthermore, a decrease in the concentration of PG490 also allows for a decrease in the absorbance of the MTS Assay, verifying that PG490 is aiding in the limiting of proliferation of the HEK cells.

However, the two drugs are not proven to be synergetic, just additive. They are not necessarily helping each other. To further investigate this, another set of graphs showing the cell viability with each drug must be plotted.
Refer to Figure 3b(ii).
From the results, another graph was compiled where the viability of the cells was compared against the concentration of IP6, without PG490. The 100% viability mark is where there was no drug used to treat the cells. The point where 50% viability is achieved, the IC50, will be noted. This graph is based on the absorbance of the HEK
cell samples. In this graph, with the cells with IP6 alone, no curve can be drawn as the points are scattered, with the majority above 100%.

Refer to Figure 3b(iii).
When the PG490, alone, is plotted against cell viability, a scatter plot is obtained with majority of the points above 100%.
Synergy is impossible to conclude as the IC50 for neither drug can be obtained as a trend line cannot be drawn.