Table 2.
Pyrosequencing results of representative samples for (A) GRHL2 and (B) RASSF1A. Pyrosequencing data validated the results from MS-HRM and provided complementary information on methylation level at each CpG position.
| A | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| % of Methylated Allele | |||||||||
|
CpG Positions
Samples |
Status by MS-HRM
(Confirmed by Pyrosequencing) |
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| 100%M Std | 90 | 88 | 89 | 86 | 90 | 83 | 96 | 94 | |
| 50%M Std | 41 | 44 | 40 | 41 | 44 | 39 | 45 | 47 | |
| 10%M Std | 12 | 16 | 15 | 14 | 16 | 16 | 18 | 19 | |
| 0%M Std | 2 | 2 | 2 | 1 | 2 | 2 | 2 | 2 | |
| BT-549 | high (yes) | 67 | 88 | 79 | 96 | 93 | 89 | 83 | 70 |
| MCF 10A | high (yes) | 74 | 77 | 72 | 88 | 89 | 76 | 79 | 53 |
| MDA-MB-231 | moderate (yes) | 17 | 27 | 15 | 25 | 20 | 23 | 21 | 13 |
| CAL-120 | very low (yes) | 6 | 12 | 7 | 7 | 9 | 8 | 8 | 5 |
| HCC1806 | 0%M (yes) | 2 | 2 | 1 | 1 | 2 | 1 | 1 | 1 |
| B | |||||||||
| % of Methylated Allele | |||||||||
|
CpG Positions
Samples |
Status by MSHRM
(Confirmed by Pyrosequencing) |
1 | 2 | 3 | 4 | ||||
| 100%M Std | 97 | 65 | 95 | 89 | |||||
| 50%M Std | 83 | 70 | 84 | 84 | |||||
| 10%M Std | 39 | 26 | 44 | 40 | |||||
| 0%M | 3 | 2 | 3 | 2 | |||||
| CAL-148 | 100%M (yes) | 89 | 92 | 96 | 96 | ||||
| SK-BR-3 | 50%M (yes) | 78 | 81 | 100 | 79 | ||||
| Hs578T | 10–50%M (yes) | 72 | 64 | 71 | 71 | ||||
| MCF 10A | moderate (yes) | 55 | 37 | 52 | 41 | ||||
| HCC1806 | 0%M (yes) | 0 | 0 | 0 | 0 | ||||
| 90–100 | 80–89 | 50–79 | 20–49 | 10–19 | 0–9 |