Data Collection:
An MTS Assay was conducted on the microtiter plate with the LNCaP 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 LNCaP data, in row B for the concentration of 0mM of IP6, the average of 0.407 and 0.695 are taken. The average of the corresponding cells in row A (0.259 and 0.216) are also averaged and then subtracted from the average of the row B results.

These values are then plotted onto the graphs Figure 4a(i) and Figure 4b(i).

Data Analysis:

Refer to Figure 4a(i).
In Figure 4a(i), there is a decrease in the absorption of the MTS Assay as the concentration of the IP6 increases, with an exception of the last column. Though this last column’s results may be a result of the formation of precipitate, the rest of the graph verifies that the IP6 is limiting the proliferation of the LNCaP cells. Furthermore, there is also a decrease in the absorbance of the MTS Assay in the LNCaP cells when the concentration of the PG490 is increase. This is showing that the PG490 is able to effectively limit the proliferation of LNCaP 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 4a(ii).
The graph compiled shows the viability of cells with increasing IP6 concentrations only. 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 cells samples. In this graph, with the cells with IP6 alone, a smooth curve line of best fit is drawn to determine the IC50, (the same trend line was achieved in the earlier experiment) which is approximately 1.3mM of IP6.

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 4a(iv).
In this graph, a descending smooth curve line of best fit was obtained, as in previous experiments. also be drawn and the IC50 can be obtained. The IC50 is determined to be 1.00ng/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 1.00ng/mL, the closest concentration to this that was used was 0.40ng/mL. The viability of the cells with 0.4ng/mL of PG490 influencing their growth is plotted against increasing concentrations of IP6.
Refer to Figure 4a(iii).
In the graph, the IC50 is determined to be 2mM, which is approximately 0.7mM off of what the value was with the IP6 drug concentrations alone. As a result, with a very little change in IC50, and no change in the trend of the graph, the PG490 does not effect the IP6 synergistically.

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 1.0mM, the closest concentration to this that was used was 1mM. The viability of the cells with 1mM of IP6 influencing their growth is plotted against increasing concentrations of PG490 in the following graph.
Refer to Figure 4a(v).
In the graph, the IC50 is determined to be 1.6ng/mL, which is only 0.6ng/mL deviated from the original IC50. As a result, with little change in IC50, the PG490 does not effect the IP6 synergistically.

Refer to Figure 4b(i).
In the second graph, there is not a clear decrease in the absorbance of the MTS Assay when the IP6 concentration is increased. In fact, there is a slight increase in absorbance. This means that the IP6 must not be working completely on these cells, as cell proliferation has not been limited. This means that IP6 must not have effect on limiting the cell proliferation of MCF7 cells. However, there is a clear decrease in the absorbance of the MTS Assay as the concentration of the PG490 is increased. This is very evident, as compared to the data when the IP6 concentration was increased. Thus, this verifies that an increase of PG490 will account of a decrease in cell proliferation.

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 4b(ii).
The graph compiled shows the viability of cells with increasing IP6 concentrations only. 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 MCF7 cells samples. In this graph, with the cells with IP6 alone, a smooth curve line of best fit is drawn to determine the IC50. However, the IC50 cannot be observed and is greater than 3.0mM of IP6.

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 4b(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 1.8ng/mL the closest concentration to this that was used was 2ng/mL.
Refer to Figure 4b(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.

The PG490 concentrations with the IC50 of IP6 cannot be graphed because an accurate IC50 for IP6 could not be determined. It is impossible to determine if the PG490 is affected by the PG490.