Fig 1: EV-SF3B4 expression and its diagnostic power in HCC. A Box plot of SF3B4 expression in blood-derived EVs from different diseases. NP, normal person; CHD, coronary heart disease; CRC, colorectal cancer; HCC, hepatocellular carcinoma; PAAD, pancreatic adenocarcinoma; WhB, whole blood; TPM, transcripts per million. B NTA of serum-derived EVs and Western blot analysis of EV and ER markers from serum EV and SNU449 cell lysates. C Gene expression of SF3B4 in MIHA and SNU449 cell lysates and derived EVs. D Representative immunofluorescence of endocytic and EV markers from MIHA cells treated with SNU449-derived EVs. E The expression of SF3B4 in MIHA cells treated with SNU449-derived EVs. F Effect of the inhibitor of clathrin-dependent endocytosis on decreasing SF3B4 expression. G SF3B4 mRNA expression comparison between healthy controls and patients with HCC in the buffy coat RNA (left), serum RNA (middle), and serum EV-RNA (right). H Expression of serum EV-SF3B4 based on the stage of liver disease in cohort 3. Statistically significant differences were determined using one-way ANOVA with Tukey’s multiple comparisons test. Compared to HC; ***P <0.001, compared to CH; ##P <0.01, ###P <0.001, compared to LC; §§§P <0.001. I Expression of serum EV-SF3B4 based on the stage of liver disease in the external cohort. Statistically significant differences were determined using one-way ANOVA with Tukey’s multiple comparisons test. Compared to HC; ***P <0.001, compared to CH; ###P <0.001, compared to LC; §§§P <0.001. J AUCs of serum EV-SF3B4 compared to serum AFP for diagnosing HCC in cohort 3. From left to right: diagnose all stages of HCC, mUICC stage I or II, and mUICC stage I. K AUCs of serum EV-SF3B4 for diagnosing HCC in the CH, LC, and HCC subgroups, compared to those of AFP in cohort 3. From left to right: diagnose all stages of HCC, mUICC stage I or II, and mUICC stage I. L AUCs of serum EV-SF3B4 compared to serum AFP for diagnosing HCC in the external cohort. From left to right: diagnose all stages of HCC, mUICC stage I or II, and mUICC stage I. M AUCs of serum EV-SF3B4 for diagnosing HCC in the CH, LC, and HCC subgroups compared to those of AFP in the external cohort. From left to right: diagnose all stages of HCC, mUICC stage I or II, and mUICC stage I
Fig 2: Single-cell RNA-seq analysis of SF3B4 enrichment in the HCC microenvironment. A UMAP plots depict cells in the human HCC microenvironment. B Expression of SF3B4 in each cell type. C Expression heatmap of the top 100 enriched genes in MDSC sub-clusters (C0–C5) (n = 504). D UMAP plot depicting MDSC subclusters. E Violin plot of the expression levels of SF3B4 in MDSC subclusters. F UMAP plot showing SF3B4+ and SF3B4- MDSCs. G Top 10 enriched terms in SF3B4 positive MDSC using the ‘MSigDB hallmark 2020’ database. H Gene Ontology depicts Biological Process (BP), Molecular Function (MF), and Cellular Component (CC) in SF3B4 positive MDSC
Fig 3: SF3B4 is overexpressed in HCC and clinically correlated with its prognosis. A SF3B4 expression in tumor and non-tumor tissues in the pan-cancer database of TCGA. B Kaplan–Meier survival analyses of SF3B4 expression in TCGA_LIHC datasets for OS, DFS, DSS, and PFS. C Forest plot of univariate Cox regression analyses of clinical parameters for OS (left) and DFS (right) (***P <0.001). D Representative images of SF3B4 expression in HCC tissues and the proportion of patients with different SF3B4 immunostaining levels in liver cancer specimens based on HPA data. Scale bar = 100 µm. E. Representative IHC photomicrographs of SF3B4 in HCC (T, tumor tissues; NT, non-tumor tissues). Scale bar = 300 µm
Fig 4: Correlation between SF3B4 and MDSC markers and their clinical relevance in TCGA LIHC. A Correlation between SF3B4 expression and tumor-infiltrating immune cells. B Correlation between SF3B4 expression and gene expression of MDSC markers. C OncoPrint of SF3B4 and its highly correlated MDSC markers in patients with HCC using TCGA_LIHC dataset ordered by baseline characteristics (overall survival status, disease-free status, neoplasm histological grade, and vascular invasion), genetic alterations, and mRNA expression levels. D Bar charts showing the proportions of alteration levels of the six gene signatures in different clinical characteristics. E Kaplan–Meier survival analyses of the altered levels of six gene signatures (6 Sig) in TCGA_LIHC datasets for OS, DFS, DSS, and PFS. F Correlation between SF3B4 expression and MDSC markers in 86 human HCC tissues
Fig 5: Efficiency of combined serum EV-SF3B4 and AFP and EV-SF3B4 positivity rates at all HCC stages in the multi-center cohort. A AUCs of the serum EV-SF3B4 and AFP combination for diagnosing HCC in the multi-center cohort. From left to right: diagnoses of all stages of HCC, mUICC stage I or II, and mUICC stage I. B AUCs of the serum EV-SF3B4 and serum AFP combination for diagnosing HCC in a subgroup composed of CH, LC, and HCC in the multi-center cohort. From left to right: diagnoses of all stages of HCC, mUICC stage I or II, and mUICC stage I. C Positive rates of AFP and serum EV-SF3B4 in patients according to liver disease status in the multi-center cohort. D The rate of positive results for serum EV-SF3B4 according to AFP status in patients with HCC, mUICC stage I or II, and mUICC stage I in the multi-center cohort
Supplier Page from MyBioSource.com for Human Splicing factor 3B subunit 4, SF3B4 ELISA Kit