A
UC TaqI
BstUI
SnaBI
C
A549
MRC5
A549
MRC5
.
BstUI
A549
MRC5
B
M
Supplemental Figure 2. A. EcSOD promoter displaying COBRA sites to determine
differential restriction between EcSOD expressing and non-expressing cell lines
after sodium bisulfite treatment. The restriction enzymes used were: SnaBI, TaqI,
and BstUI.
B. Cobra results for TaqI, BstUI and SnaBI in the EcSOD-positive
MRC-5 cells versus EcSOD-negative A549 cells. Enzymes cut when when the cytosine in the
enzymes cut site remained methylated and unconverted by sodium bisulfite. These results
confirm that the EcSOD promoter is aberrantly hypermethylated in an EcSOD-negative cells.
C. COBRA results of BstUI restriction in the clinical lung adenocarcinoma tumors showing
a greater methylation pattern in 4 of the 5 tumor samples when compared to normal lungown ,
These results further confirm results shown in the heatmap in Figure 3 that downregulation of
EcSOD in lung tumors is associated with a more hypermethylated EcSOD promoter.