Fig 2: Serpinc1 overexpression induced apoptosis in HepG2 and SMMC7721 cells. (A–F) HepG2 and SMMC7721 cells were overexpressed with control or serpinc1. (A) Western blot to detect the expression level of serpinc1. (B) MTS to detect the cell viability. (C, D) Detection of cell cycle distribution. Representative images (C) and statistical analysis (D) of cell cycle distribution. (E) Wound healing assay to analyze the migration of hepatocellular carcinoma cell lines. (F) Flow cytometry to analyze the apoptosis. (G, H) Scrambled or serpinc1 shRNA were transfected in HepG2 and SMMC7721 cells. The knockdown of serpinc1 was detected by western blot (G), and the cell apoptosis was detected by flow cytometry (H). *P < 0.05, ns, no significant, P > 0.05.

Fig 3: Network of differentially expressed proteins in KD with CAD and CAA. (A) Network based on functionally-enriched KEGG pathways. The proteins in the network were differentially expressed proteins in the present study. (B) Network associated with the variations within patients with KD and CAD/CAA. CFH (yellow cluster), MBL2 (pink cluster), KNG1 (grey cluster), FN1 (green cluster) and SERPINC1 (green cluster) were identified as primary candidates involved in CAA and CAD (presented as bigger nodes with red circles). P<0.05 was a threshold for KEGG pathway enrichment. KD, Kawasaki disease; CAA, coronary artery aneurism; CAD, coronary artery dilation; KEGG, Kyoto Encyclopedia of Genes and Genomes; FN1, fibronectin 1; CFH, complement factor; KNG1, kininogen 1; MBL2, mannose binding lectin 2; SERPINC1, serpin family C member 1; FDR, false discovery rate.

Fig 4: Validation of proteomics results by western blot analysis. (A) Western blot analysis was used for the validation of 5 proteins (KRT1, APOB, FN1, SERPINC1 and SERPINA1) from control, -Treatment and +Treatment samples; ß-actin was used as an internal control. (B) Histogram of western blot analysis. The control group is shown in gray, the -Treatment group is blue and the +Treatment group is brown. Data are expressed as mean ± standard deviation (n=25 biological replicates in each group). The multiple comparisons test of control vs. -Treatment and control vs. +Treatment are performed using analysis of variance with Dunnett's test. *P<0.05, **P<0.01 and ***P<0.001 vs. control. KRT1, keratin 1; APOB, apolipoprotein B; FN1, fibronectin 1; SERPINC1, antithrombin III; SERPINA1, a-1-antitrypsin.

Fig 5: Serpinc1 protein expression is correlated with outcomes in hepatocellular carcinoma patients. (A–D) Analysis of the correlation between the serpinc1 expression and survival rate in hepatocellular carcinoma patients from Kaplan-Meier Plotter database. Overall survival (A), Progression-free survival (B), Relapse-Free survival (C), and Disease-free survival (D). (E, F) Analysis of the effect of alcohol and serpinc1 expression on the outcomes from Kaplan-Meier Plotter database. Alcohol (E) and none alcohol (F). (G, H) Detection of the effect of Hepatitis virus and serpinc1 expression on the outcomes from Kaplan-Meier Plotter database. Hepatitis virus (G) and none hepatitis virus (H). (I–L) Analysis of the effect of serpinc1 expression and sorafenib administration on outcomes of hepatocellular carcinoma patients from Kaplan-Meier Plotter database. Overall survival (I), Progression-free survival (J), Relapse-Free survival (K), and Disease-free survival (L).