Basic, Underlying Data for Figures 3-5

Basic, Underlying Data for Figures 3-5

Supplemental Figure 1a. GC-Rich Regions of Altered Methylation: Comparison of DMBA-Initiated, 3mg CSC-promoted (8wks) Tissue to Control (DMBA/Acetone) % DMBA/Acetone Control PCR Product Size (bp)

Supplemental Figure 1b. GC-Rich Regions of Altered Methylation (New Methylations Only): Comparison of DMBA-Initiated, 3mg CSC-promoted (8wks) Tissue to Control (DMBA/Acetone) Peak Area (Peak Area Units) PCR Product Size (bp)

Legend - Supplemental Figure 1.GC-Rich Regions of Altered Methylation: Comparison of DMBA-Initiated, 3mg CSC-promoted (8wks) Tissue to Control (DMBA/Acetone). RsaI/HpaII (blue symbols) and RsaI/MspI (red symbols) digestion and subsequent AP-PCR was performed on DNA isolated from SENCAR mouse skin treated with 3mg CSC for 8wks. Regions of hypomethylation were prevalent (a) Regions of new methylation resulting from treatment are shown in terms of the peak area for each PCR product size (b). A table tallying the regions of altered methylation for each treatment are shown as an inset in each figure. Regions of hypo-, hyper-, and new methylation are expressed in terms of the treated mean for each PCR product size as a percent of the control mean for each PCR product size. All changes projecting below the x-axis represent decreases in methylation (hypomethylation) while all those above the x-axis represent increases in methylation (hypermethylation). All 100% hypomethylations are considered to be significant, and only the hypermethylations and partial hypomethylations that were statistically significantly different from control values (Student’s t-test, p<0.05) are depicted.

Legend - Supplemental Figure 2.GC-Rich Regions of Altered Methylation: Comparison of DMBA-Initiated, 9mg CSC-promoted (8wks) Tissue to Control (DMBA/Acetone). RsaI/HpaII (blue symbols) and RsaI/MspI (red symbols) digestion and subsequent AP-PCR was performed on DNA isolated from SENCAR mouse skin treated with 9mg CSC for 8wks. Regions of hypomethylation and hypermethylation are shown (a). Regions of new methylation resulting from treatment are shown in terms of the peak area for each PCR product size (b). A table tallying the regions of altered methylation for each treatment are shown as an inset in each figure. Regions of hypo-, hyper-, and new methylation are expressed in terms of the treated mean for each PCR product size as a percent of the control mean for each PCR product size. All changes projecting below the x-axis represent decreases in methylation (hypomethylation) while all those above the x-axis represent increases in methylation (hypermethylation). All 100% hypomethylations are considered to be significant, and only the hypermethylations and partial hypomethylations that were statistically significantly different from control values (Student’s t-test, p<0.05) are depicted.

Supplemental Figure 3c. GC-Rich Regions of Altered Methylation (New Methylations Only): Comparison of DMBA-Initiated, 18mg CSC-promoted (8wks) Tissue to Control (DMBA/Acetone) 151236 285828 143137 278758 Peak Area (Peak Area Units) PCR Product Size (bp)

Legend - Supplemental Figure 3.GC-Rich Regions of Altered Methylation: Comparison of DMBA-Initiated, 18mg CSC-promoted (8wks) Tissue to Control (DMBA/Acetone). RsaI/HpaII (blue symbols) and RsaI/MspI (red symbols) digestion and subsequent AP-PCR was performed on DNA isolated from SENCAR mouse skin treated with 18mg CSC for 8wks. Regions of hypomethylation and hypermethylation are shown (a). Regions of new methylation resulting from treatment are shown in terms of the peak area for each PCR product size (b). The y-axis scale was expanded in order to facilitate visualization of all data points. In doing this four regions of new methylation whose peak areas exceeded the scale of the chart were labeled above the chart with their corresponding peak area values (c). A table tallying the regions of altered methylation for each treatment are shown as an inset in each figure. Regions of hypo-, hyper-, and new methylation are expressed in terms of the treated mean for each PCR product size as a percent of the control mean for each PCR product size. All changes projecting below the x-axis represent decreases in methylation (hypomethylation) while all those above the x-axis represent increases in methylation (hypermethylation). All 100% hypomethylations are considered to be significant, and only the hypermethylations and partial hypomethylations that were statistically significantly different from control values (Student’s t-test, p<0.05) are depicted.

Supplemental Figure 6a. GC-Rich Regions of Altered Methylation: Comparison of Tumor Tissue to Control (DMBA/Acetone) % DMBA/Acetone Control PCR Product Size (bp)

Supplemental Figure 6b. GC-Rich Regions of Altered Methylation (New Methylations Only): Comparison of Tumor Tissue to Control (DMBA/Acetone) Peak Area (Peak Area Units) PCR Product Size (bp)

Legend - Supplemental Figure 6.GC-Rich Regions of Altered Methylation: Comparison of Tumor Tissue to Control (DMBA/Acetone). RsaI/HpaII (blue symbols) and RsaI/MspI (red symbols) digestion and subsequent AP-PCR was performed on DNA isolated from tumors resulting from treatment with CSC. Regions of hypomethylation and hypermethylation are shown (a). Regions of new methylation identified are shown in terms of the peak area for each PCR product size (b). A table tallying the regions of altered methylation for each treatment are shown as an inset in each figure. Regions of hypo-, hyper-, and new methylation are expressed in terms of the treated mean for each PCR product size as a percent of the control mean for each PCR product size. All changes projecting below the x-axis represent decreases in methylation (hypomethylation) while all those above the x-axis represent increases in methylation (hypermethylation). All 100% hypomethylations are considered to be significant, and only the hypermethylations and partial hypomethylations that were statistically significantly different from control values (Student’s t-test, p<0.05) are depicted.

Basic, Underlying Data for Figures 3-5