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Supplementary Table 1. Clinicopathologic Information for Cases MBC 1-10

Case Post-Mortem Interval

Age at Diagnosis

Stage at Diagnosis

Primary Tumor Type and Grade

Age at Death

1 3 hours, 45 minutes

54 T2N0M0 ILC, grade 2 65

2 4 hours 33 T3N1M0 IDC, grade 3 37 3 2 hours 40 T1N1M0 IDC, grade 2 48 4 3 hours 44 T1miN0M0 IDC, grade 3 49 5 4 hours 59 T1N0M0 IDC, grade 2 68 6 3 hours, 30

minutes 56 T2N1M0 IDC, grade 2 61

7 1 hour, 15 minutes

51 T2N0M0 IDC, grade 3 53

8 3 hours 48 T2N3M0 IDC, grade 3 54 9 3 hours 38 T2NxM0 IDC, grade 3 43 10 3 hours 71 T2N0M0 IDC, grade 3 79

Post-Mortem Interval

Age at Diagnosis

Stage at Diagnosis

Primary Tumor Type and Grade

Age at Death

3 hours, 45 minutes

54 T2N0M0 ILC, grade 2 65

4 hours 33 T3N1M0 IDC, grade 3 37 2 hours 40 T1N1M0 IDC, grade 2 48 3 hours 44 T1miN0M0 IDC, grade 3 49 4 hours 59 T1N0M0 IDC, grade 2 68 3 hours, 30

minutes 56 T2N1M0 IDC, grade 2 61

1 hour, 15 minutes

51 T2N0M0 IDC, grade 3 53

3 hours 48 T2N3M0 IDC, grade 3 54 3 hours 38 T2NxM0 IDC, grade 3 43 3 hours 71 T2N0M0 IDC, grade 3 79

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Supplementary Table 2: Sites of Metastasis.

Case Liver Bone Lung Brain Adrenal GI Tract Mesentery Other MBC1 Extensive Rib +minimal - + Left Pancreas,

gallbladder Extensive

MBC2 + spine Extensive + Bilateral - + Axillary LN, ovary

MBC3 Extensive Spine + + + Right - + Left ovary MBC4 Extensive Spine + + Bilateral Small bowel + Spleen MBC5 Extensive Spine + + +Right Pancreas,

stomach _ Fallopian tube,

breast, pericardium, pleura

MBC6 Extensive Spine + - - Stomach, cecum

- Pleura, pericardium

MBC7 Extensive Spine - + Bilateral - + Mediastinal, axillary LN

MBC8 Extensive Spine +minimal - - - - Diaphragm MBC9 - Spine,

femur + NE - Pancreas - Abdominal

LN, pleura MBC10 + NE + - +Left - + Mediastinum,

pericardium, omentum, diaphragm

NE = not examined at autopsy Bold = extensive LN=lymph node GI=gastrointestinal

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Supplementary Table 3: Mesothelin Immunohistochemistry Case Primary Tumor Positive Metastases Negative Metastases MBC 1 Negative None Liver, Bone,

Mesentery, Gallbladder, Rectum

MBC 2 1+ Bone, Axillary LN, Brain (1+)

Lung, Adrenal, Liver, Abdominal LN, Ovary

MBC 3 Negative None Bone, Liver, Ovary, Brain, Mesentery, Lung

MBC 4 N/A Lung Liver, Pericardium, Spleen, Small Bowel, Adrenal, Bone

MBC 5 Negative None Lung, Liver, Pancreas, Fallopian Tube, Adrenal, Brain, Pericardium, Bone, Pleura

MBC 6 Negative Pleura, Lung, Colon (1+)

Liver, Axillary Lymph Node, Bone

MBC 7 Positive (2+) Omentum, Mediastinal LN(1+)

Diaphragm, Axillary Metastases, Adrenal, Bone, Liver

MBC 8 Negative None Pleura, Liver, Bone, Axillary LN

MBC 9 Negative None Bone, Abdominal LN, Adrenal

MBC 10 Negative None Lung, Hilar LN, Liver, Pericardium, Adrenal, Omentum

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Supplementary Table 4: Cox-2 Immunohistochemistry Case Primary Tumor Positive Metastases Negative Metastases MBC 1 Negative Liver, Mesentery

(2+), Bone (3+) Gallbladder, Rectum

MBC 2 1+ Left Adrenal Gland , Bone, Liver, Abdominal LN, Lung, Ovary, Diaphragm (1+); Right Adrenal Gland, Axillary LN (2+)

Brain

MBC 3 Negative None Bone, Liver, Ovary, Brain, Mesentery, Lung

MBC 4 N/A Liver, Adrenal (1+) Lung, Pericardium, Spleen, Small Bowel, Bone

MBC 5 Negative None Lung, Liver, Pancreas, Fallopian Tube, Adrenal, Brain, Pericardium, Bone, Pleura

MBC 6 Negative Axillary Lymph Node, Liver, Pleura, Lung, Bone (1+)

Colon

MBC 7 Negative Omentum, Mediastinal LN, Diaphragm, Right Adrenal (1+)

Liver, Left Adrenal, Bone

MBC 8 Negative Bone (1+) Pleura, Liver, Axillary LN

MBC 9 Positive (2+) None Bone, Abdominal LN, Adrenal

MBC 10 Positive (2+) Liver, Mediastinal LN, Adrenal, Lung (1+); Hilar LN (2+)

Omentum, Pericardium

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Supplementary Table 5 : Percentage Promoter Methylation of RASSF1A, HIN1, CYCLIN D2, TWIST, RARβ, APC1, and ERα in lesions of primary and metastatic breast cancer. Scores for cumulative methylation were obtained by QM-MSP assay in liver , mesentery, gallbladder, bone, lung (lobes of right upper = RUL, right middle = RML, right lower = RLL, and left lower = LLL), ovary , adrenal , kidney, rib soft tissue, pericardium, omentum, diaphragm, and lymph node [abdominal (Abd) and mediastinal (Med)]. The % methylation = 100 X [ # copies methylated DNA / (# copies unmethylated + methylated DNA)] contained an aliquot produced from multiplex methylation-independent PCR wherein the entire gene panel was co-amplified before performing quantitative real-time MSP analyses of each gene.

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Tissue RASSF1A HIN1 CYCLIN D2 TWIST RAR B APC1ER

ALPHAMBC001 Primary 6.840 0.000 0.000 na 0.000 43.179 0.000

Liver #1 11.352 0.000 2.924 18.483 4.422 28.052 0.000Liver #4 24.005 0.000 13.767 53.007 3.454 77.666 0.000Mesentery 24.047 0.000 60.268 86.402 32.348 85.684 0.091Mesentery 26.789 0.065 36.007 75.656 28.135 79.247 0.032Gall Bladder 26.882 0.987 30.472 68.654 28.884 65.913 0.051Bone 23.029 0.000 46.280 92.201 0.000 99.635 0.000

MBC002 Primary 0.000 0.133 7.117 13.621 2.447 0.068 0.003Lung, RML 0.027 0.040 0.000 72.768 1.402 1.889 0.000Lung, RUL 0.000 0.000 0.000 62.985 2.342 1.131 0.000Ovary 0.000 0.000 0.000 77.819 0.758 0.000 0.000Lung, RLL 0.000 0.011 0.001 72.597 0.955 1.218 0.000Adrenal 0.003 0.000 0.000 55.028 1.335 0.000 0.000Kidney 0.000 0.000 0.000 69.925 3.191 0.001 0.000Bone 0.002 0.018 0.000 45.917 0.128 0.000 0.013

MBC003 Primary 77.900 99.956 93.700 65.260 1.437 0.236 0.000Liver #1 92.768 90.452 83.054 0.174 1.083 0.000 0.001Liver #1 87.723 96.594 77.529 0.074 1.505 2.109 naSoft tissue rib 85.441 88.431 71.988 0.176 1.109 0.000 0.009Liver #2 87.280 79.002 83.094 0.052 0.615 2.377 2.047Lung, RLL 40.319 27.412 51.609 1.563 0.881 1.725 0.529Bone 13.956 93.364 47.507 0.001 0.002 0.504 0.828

MBC004 Adrenal 1.461 0.028 0.000 0.010 0.090 29.379 0.0000Pericardium 0.000 0.501 0.004 4.445 3.929 22.172 0.0780Liver #2 1.805 6.158 0.076 1.164 0.162 51.272 0.1036Liver #3 0.168 12.500 2.184 0.952 0.015 70.035 1.4547Lung, RLL 0.000 15.081 0.062 2.190 0.369 32.282 0.0018Lung, LLL 0.801 5.336 1.469 1.754 0.149 17.356 0.0000

MBC005 Primary 48.562 90.226 29.316 0.000 4.961 37.314 0.000Lung, RUL 51.396 83.641 57.760 0.144 4.305 51.029 0.034Pericardium 75.776 91.840 78.016 0.044 4.711 23.871 0.000Liver, Left 66.513 89.146 77.099 0.207 5.140 0.447 0.429Omentum 92.789 99.805 42.152 0.022 8.410 1.792 0.000Liver, Right 74.196 90.749 76.298 0.012 4.274 0.884 1.531Pancreas 60.214 75.503 66.476 0.117 5.765 0.749 0.737

MBC006 Primary 31.838 0.000 0.845 0.041 6.519 0.804 0.053Liver, Right 1 25.977 0.189 2.135 0.058 10.062 27.669 1.953Liver, Right 2 49.603 0.003 3.424 0.017 8.819 26.412 2.468Liver, Left 28.579 0.000 10.492 0.049 9.802 10.370 0.503Bone 0.032 0.078 0.000 0.098 0.032 0.000 0.000

MBC007 Primary 66.707 0.000 0.000 0.000 7.653 6.196 0.000Omentum 69.271 0.000 6.283 0.098 6.778 0.000 0.080Diaphragm 94.964 0.000 11.253 0.011 11.598 0.000 0.000Liver, #1 30.161 0.000 11.832 0.395 12.710 34.493 3.830Liver, #4 72.062 0.000 7.199 0.003 12.903 14.270 1.609Adrenal 89.345 0.000 1.329 0.004 8.167 0.000 0.000Bone 5.243 0.048 1.748 1.323 0.094 0.014 0.045

MBC008 Primary 64.470 100.000 45.061 48.206 14.680 0.000 0.000Liver, Left #1 74.425 95.905 78.603 79.092 3.871 0.847 0.141Liver, Right #1 86.350 99.975 91.810 95.276 8.232 1.812 0.790Liver, Right # 2 72.102 98.485 70.680 72.958 4.528 2.084 0.002Liver, Left #2 90.330 99.831 97.075 85.864 9.632 0.018 0.003

MBC009 Primary 53.572 69.797 29.090 58.917 4.894 0.900 0.0210Adrenal 70.785 84.057 0.017 57.226 0.760 2.378 0.0004Lung, RML 77.269 85.965 0.033 80.960 0.736 22.904 0.0000Lymph node, Abd #1 78.182 76.728 0.000 69.305 0.520 25.000 0.0000Lymph node, Abd #3 69.385 68.310 0.012 69.698 0.214 24.387 0.0000Lung, RLL 40.298 43.007 0.005 50.759 0.439 25.171 0.0000Lung, LUL 4.936 4.864 0.081 14.173 1.349 5.956 0.0000

MBC010 Primary 3.051 0.000 83.827 58.212 10.772 75.851 0.0000Omentum 0.264 0.000 76.811 81.418 0.546 50.592 0.0000Adrenal 1.110 0.000 60.031 50.048 0.531 27.777 0.0486Lymph node, Med #1 2.859 13.451 74.165 75.244 0.218 43.717 0.0000Lymph node, Med #5 1.892 0.000 69.487 76.638 0.639 32.946 0.0000Lung, RLL 0.103 0.471 43.110 58.325 1.532 24.474 0.0000Lung, LLL 0.000 0.063 61.754 66.961 5.267 74.767 0.0216Liver 0.649 0.147 64.533 34.632 0.228 43.530 1.8245