Additional file 7 of LncRNA FAM83H-AS1 promotes the malignant progression of pancreatic ductal adenocarcinoma by stabilizing FAM83H mRNA to protect β-catenin from degradation

Additional file 7: Supplementary Figure 1. Hierarchical cluster heat map of differentially expressed lncRNAs in PDAC and corresponding normal tissues generated from RNA sequencing data from the TCGA database. Red in the heat map represents upregulation; Green represents downregulation. 817 differentially expressed lncRNAs were screened out in the comparing mode between PDAC and pancreas. Supplementary Figure 2. Hierarchical cluster heat map of differentially expressed lncRNAs in T2N0M0 PDAC and T2N1M0/T2N0M1/T2N1M1 PDAC tissues generated from RNA sequencing data from the TCGA database. Red in the heat map represents upregulation; Green represents downregulation. 275 differentially expressed lncRNAs were screened out in the comparing mode between T2N0M0 and T2N1M0/T2N0M1/T2N1M1. Supplementary Figure 3. Hierarchical cluster heat map of differentially expressed lncRNAs in T3N0M0 PDAC and T3N1M0/T3N0M1/T3N1M1 PDAC tissues generated from RNA sequencing data from the TCGA database. Red in the heat map represents upregulation; Green represents downregulation. 169 differentially expressed lncRNAs were screened out in the comparing mode between T3N0M0 with T3N1M0/T3N0M1/T3N1M1. Supplementary Figure 4. Survival analysis was used to illustrate the relationship between the level of 6 lncRNAs (FAM83H-AS1, LINC00365, LINC00628, LINC00261, AFAP1-AS1, and HNF1A-AS1) and overall survival in TCGA cohort, respectively. Supplementary Figure 5. Migration capacities of 6 lncRNAs were estimated in vitro. (A) Transwell assays were used to determine the migration capabilities of 6 lncRNAs siRNA-transfected PDAC cells, respectively; Scale bar: 100 μm. (B) Wound healing assays were conducted to evaluate the migration abilities of 6 lncRNAs siRNA-transfected PDAC cells, respectively; Scale bar: 100 μm. *P < 0.05, **P < 0.01, ***P < 0.001. Supplementary Figure 6. Effect of knockdown of 6 candidate lncRNAs on the filamentous state of F-actin. SW 1990 cells were treated with lncRNAs siRNA or control siRNA respectively and analyzed by immunofluorescence, which were estimated the bundling and disassembling sates of F-actin. Supplementary Figure 7. Expression of FAM83H-AS1 in serums were tested in the patients with PDAC, benign non-pancreatic disease and benign pancreatic disease from Tongji hospital. Supplementary Figure 8. Survival analysis was used to illustrate the relationship between the level of FAM83H-AS1 and overall survival in high purity TCGA cohort, in which the samples below 60% neoplastic cellularity were excluded. Supplementary Figure 9. CCK-8 assays were used to compare the cell viability between LV-FAM83H-AS1 group and LV-Control group in PANC-1 and SW 1990 cells. ***P < 0.001. Supplementary Figure 10. FAM83H expression in stable FAM83H overexpressed and down-expressed PDAC cells, confirmed by RT-qPCR analysis. *P < 0.05, **P < 0.01, ***P < 0.001. Supplementary Figure 11. FAM83H-AS1 promotes β-catenin nuclear location via FAM83H. Western blot for FAM83H, β-catenin, GAPDH, and Histone-H3 with the protein lysates fractionated according to subcellular localization after overexpressing FAM83H-AS1 and knocking down FAM83H in PANC-1 and SW 1990 cells