Supplemental Figures. Amylase. Glucose. Pancreas FAP-WT FAP-KO weight (g) mg/dl U/L 0.

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1 Supplemental Figures A U/L 4 mg/dl weight (g) Amylase Glucose Pancreas B Trichrome HABP PAS H&E Supplemental Figure 1. FAP is dispensable for pancreatic development. (A) The pancreas was harvested from and C57BL/6 mice and weighed. Serum glucose and amylase were measured. Results are shown as mean ± SEM (n=6). (B) Representative hematoxylin and eosin (H&E), trichrome, hyaluronan-binding peptide (HABP) and periodic acid Schiff (PAS)-stained pancreas from and C57BL/ 6 mice; scale: 1 µm. No significant difference was found.

Expression of FAP is haplosufficient and its genetic deletion does not result in stromal cell ablation in pancreatic tumors.

2 A B CD45 CD9+ cells Mouse adult foreskin fibroblasts FAP-Het 48 FAP FAP C vimentin 44.2 FAP CD9 CD9 CD9 % of Max FAP-Het.369 Sheep-IgG isotype vimentin isotype % of area Supplemental Figure 2. Expression of FAP is haplosufficient and its genetic deletion does not result in stromal cell ablation in pancreatic tumors. (A) Analysis of FAP expression in mouse adult fibroblasts isolated from FAP-wild type (FAPWT), FAP-heterozygous (FAP-Het) or FAP-knockout () C57BL/6 mice. and FAP-Het fibroblasts exhibited similar FAP expression. (B) Analysis of FAP expression in orthotopically implanted PanO2 PDA cells in or FAP-Het C57BL/6 mice. Intratumoral CD45 CD9+ cells from and FAP-Het mice showed equivalent FAP expression. (C) Syngeneic KPC tumor cells were implanted subcutaneously in and mice and harvested for vimentin staining to evaluate stromal cell content. scale: 5 µm. Results are shown as mean ± SEM (n=3). No significant difference was found.

3 A B Tumor volume (mm 3 ) FH-KPC FKO-KPC Tumor number FH-KPC FKO-KPC C D PDA subtype (% of total mice) FH-KPC (n=27) FKO-KPC (n=23) % of mice with glandular PDA FH-KPC (n=21) FKO-KPC (n=16) Glandular Sarcomatoid Anaplastic Well Moderate Poor Differentiated Undifferentiated Differentiation status Supplemental Figure 3. Targeting FAP does not impact the size, number, subtypes or differentiation of pancreatic tumors in the KPC mouse model. (A) FH-KPC and FKO-KPC mice with PDAs were digitally visualized and reconstructed to determine tumor volumes using the Integrated Vevo Workstation software package. (B) The number of PDAs from FH-KPC and FKO-KPC mice was assessed during ultrasound scanning. Data shown in (A) and (B) represent mean ± SEM (FH-KPC, n=25; FKO-KPC, n=23). No statistical significance was found in tumor volume or tumor number by Student s t- test. (C) PDAs derived from FH-KPC and FKO-KPC mice exhibited a mix of histological phenotypes (subtypes): glandular type, sarcomatoid type and anaplastic type. The predominant histology of each tumor was defined by the presence of more than 5% of a subtype within each sample as determined by a board-certified pathologist (ELB). (D) The differentiation status of the glandular PDA was further evaluated and scored. No statistical significance was found by chi-square test in tumor subtypes or differentiation.

4 Supplemental Table 1. Summary of clinicopathological features of primary pancreatic ductal carcinoma from 121 patients analyzed for FAP expression. Characteristics Patient number (%) Age (years) < > Sex Male Female Alcohol No Yes Smoking No Yes pt(primary tumor) pt pt pt pt pn(regional lymph node metastasis) pn pn pm(distant lymph node metastasis) pm pm1 1.8 Recurrence No Yes Microvessel invasion No Yes 1 8.3

5 Supplemental Table 2. Relationship between FAP expression and clinicopathological factors in primary pancreatic ductal carcinomas. Characteristics Low (, 1) n=28 FAP expression High (2, 3) n=93 Age Years (mean ± SD) 67.3 ± ± 12.2 Sex Male Female 9 3 Alcohol status No Yes 7 28 Smoking status No Yes 5 34 Tumor size Centimeter (mean ± SD) 3.4 ± ± 1.43 Stage I 4 9 II III 7 IV 1 I + II III + IV 1 7 Tumor status T1 1 2 T T T4 7 P-value.735 a 1. b.613 b.634 b.7979 a.1189 b.462 b.4278 b.3643 b T1 + T T3 + T Lymph node status 1. b N 12 4 N Distal metastasis status.2314 b M M1 1 Recurrence status - No Yes Microvessel invasion status.5913 b No Yes 3 7

6 Student s t test and Pearson chi-square test were performed for continuous variables and categorical variables, respectively. The tumor stage, tumor, lymph node, and distal metastasis status were classified according to the international system for staging pancreatic cancer. SD represents standard deviation. *P value <.5 was considered statistically significant. a denotes Student s t test b Chi-square test

7 Supplemental Table 3. COX univariate regression and COX multiple regression analysis of prognostic factors for overall and disease-free survival in 121 patients with pancreatic ductal carcinoma. Cox univariate regression (Overall survival) Variables Comparison Hazard Ratio (95% CI) P-value T T2-4; T ( ).451 N N1; N 1.88 ( ).649 M M1; M ( ).2 * FAP High (2,3); Low (,1) ( ).7 * Cox multiple regression (Overall survival) Variables Comparison HR (95% CI) P-value T T2-4; T ( ).573 N N1; N 1.13 ( ).943 M M1; M 5. ( ).1 * FAP High (2,3); Low (,1) ( ).5 * Cox univariate regression (Disease-free survival) Variables Comparison HR (95% CI) P-value T T2-4; T ( ).22 N N1; N ( ).464 M M1; M ( ).13 FAP High (2,3); Low (,1) 3.21 ( ) <.1 * Cox multiple regression (Disease-free survival) Variables Comparison HR (95% CI) P-value T T2-4; T ( ).291 N N1; N 1.58 ( ).766 M M1; M ( ).18 * FAP High (2,3); Low (,1) ( ) <.1 *

Supplementary Information

Supplementary Information Prognostic Impact of Signet Ring Cell Type in Node Negative Gastric Cancer Pengfei Kong1,4,Ruiyan Wu1,Chenlu Yang1,3,Jianjun Liu1,2,Shangxiang Chen1,2, Xuechao Liu1,2, Minting