Figure 2. State of Cell Types Without Treatment (Control Only)

A) Raman spectra from wavenumbers 2600 cm^-1 to 3200 cm^-1 of lipid droplets in MB-MDA-231 cells. At the 2850 cm^-1 lipid wavenumber, there is higher peak intensity in the 20x methionine treatment compared to the 1x condition. At the 2928 cm^-1 protein wavenumber, there is higher peak intensity in the 1x methionine condition compared to the 20x methionine.

B) Comparison of C-D Lipid/ Lipid peak ratios across cell types. Ratios of C-D (carbon-deuterium labeled) lipid to total lipid content were quantified using their Raman wavenumbers (2140 and 2850 cm^-1 respectively). While MDA231 and MCF7 had comparable CD labeled lipid- lipid ratios (differences report p>0.05), they both were higher than MCF10A (p<0.05).

Figure 3. Effects of Methionine Within and Across Cell Types

A) Comparison of Cholesterol/Lipid peak ratios within cell types for control and 20x methionine conditions. Ratios of cholesterol to total lipid content were quantified using their Raman wavenumbers (2883 and 2850 cm^-1 respectively). Excess methionine did not significantly alter cholesterol-lipid ratios in MCF10A and MCF7 cells (differences report p>0.05), however excess methionine resulted in a lower cholesterol-lipid ratio compared to normal conditions in MDA231 (p<0.05).

B) Comparison of effects of methionine across cell types. Differences in peak ratios after methionine treatment are plotted for each cell type. The directionality of the effect of excess methionine yields a significant difference (p<0.05) for cancer cells.

Figure 4. Effects of Insulin Within and Across Cell Types

A) Comparison of Lipid/Protein peak ratios across cell types. Ratios of lipid to protein content were quantified using their Raman wavenumbers (2850 and 2928 cm^-1 respectively). In a comparison of controls, MCF10A had the highest lipid-protein ratio, with MCF7 having the second most, and MDA231 having the least (all p<0.05).

B) Comparison of effects of increased insulin (2x) within and across cell types. In terms of directionality of change comparing excess insulin to their controls, the cancer cells demonstrated an increase in lipid/protein ratio (differences report p>0.05) while the MCF10A cells demonstrated a decrease in lipid/protein ratio (p<0.05). Though the control groups show a significant difference in ratios prior to treatment, post-treatment ratios were comparable for MCF7 and MDA231 (differences report p>0.05) but still were significantly different than MCF10A (p<0.05 for both).

Figure 5. Effects of Methionine and Insulin Within and Across Cell Types

A) Effects of excess concentration of metabolites within cell types. Comparing increased insulin and increased methionine levels to normal conditions, MDA231 demonstrated an increased lipid/protein ratio (p<0.05), while the effect was negligible in MCF7 and MCF10A (differences report p>0.05).

B) Effects of excess concentration of metabolites across cell types. In terms of directionality of change due to excess metabolites compared to control, MCF10 and MCF7 resulted in a lower lipid/protein ratio (differences report p>0.05). However, excess metabolites resulted in an increased lipid protein ratio for MDA231 (p<0.05). The differences between MDA231 and the other cell types were significant (p<0.05).

Made by Summer Joyce Batasin