Table S2 Study group sample sizes for CEA, CYFRA21-1 and CA125 determinations.

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1 Supplementary Data Table S Clinico-pathological data associated with malignant and benign cases Primary site Early stage Late stage Caecum 3 (5%) 4 (20%) (5%) Ascending colon 6 (30%) 2 (0%) 0 (0%) Transverse colon 0 (0%) 3 (5%) 3 (5%) Splenic flexure (5%) 0 (0%) 0 (0%) Descending colon (5%) 0 (0%) 2 (0%) Sigmoid colon 5 (25%) 5 (25%) 8 (40%) Rectosigmoid 0 (0%) 0 (0%) (5%) Rectum 3 (5%) 6 (30%) 2 (0%) Undetermined (5%) 0 (0%) 3 (5%) Standardised stage A 7 (35%) 0 (0%) NA B 3 (65%) 0 (0%) NA C 0 (0%) 4 (70%) NA D 0 (0%) 6 (30%) NA Grade I (5%) 2 (0%) NA II 2 (60%) 2 (60%) NA III 3 (5%) 2 (0%) NA Unknown 4 (20%) 4 (20%) Morphology Adenocarcinoma 20 (00%) 20 (00%) NA Table S2 Study group sample sizes for CEA, CYFRA2- and CA25 determinations. CEA and CYFRA2-/CA25 Years to diagnosis Early control Early case Late control Late case 0-20/24 20/23 8/20 20/30 20/29-2 9/24 8/23 20/20 8/2 9/ /20 9/23 8/24 7/22 7/ /20 8/20 20/22 20/22 20/27

2 All cases vs. + Table S3 Tumour marker performance according to different cut-off thresholds (ng/ml) and pre-diagnosis time group. CEA or CYFRA refers to a combined panel where an elevated level of either marker is considered positive. (A) Cancer cases versus non-cancer controls + benigns; (B) Cancer cases versus benign only. Green shading for sensitivity and red shading for specificity is scaled from highest to lowest value in each table. (A) Time group 0- yr CEA CYFRA CEA or CEA CYFRA CEA or CEA CYFRA CEA or CEA CYFRA CEA or Number/Parameter >5 ng/ml >3.3 ng/ml CYFRA >3 ng/ml >2 ng/ml CYFRA >2.5 ng/ml >.5 ng/ml CYFRA >2.5 ng/ml >3.3 ng/ml CYFRA positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%)

3 All s vs. (B) Time group 0- yr CEA CYFRA CEA or CEA CYFRA CEA or CEA CYFRA CEA or CEA CYFRA CEA or Number/Parameter >5 ng/ml >3.3 ng/ml CYFRA >3 ng/ml >2 ng/ml CYFRA >2.5 ng/ml >.5 ng/ml CYFRA >2.5 ng/ml >3.3 ng/ml CYFRA positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%) yr positive (n) negative (n) positive (n) negative (n) Sensitivity (%) Specificity (%)

4 (A) Early stage cases Figure S Serum CEA longitudinal profiles in individual case control samples. (A) Early stage cases; (B) Late stage cases. Horizontal line represents 5 ng/ml CEA. Early stage Early stage 2 Early stage 3 Early stage Early stage 5 Early stage 6 Early stage 7 Early stage Early stage 9 Early stage 0 Early stage Early stage Early stage 3 Early stage 4 Early stage 5 Early stage Early stage 7 Early stage 8 Early stage 9 Early stage

5 (B) Late stage cases Late stage Late stage 2 Late stage 3 Late stage Late stage 5 Late stage 6 Late stage 7 Late stage Late stage 9 Late stage 0 Late stage Late stage Late stage 3 Late stage 4 Late stage 5 Late stage Late stage 7 Late stage 8 Late stage 9 Late stage

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