Journal: Journal of Hepatocellular Carcinoma

Article Title: Decoding ARF4 and EIF5B-Based Prognostic Signatures and Immune Landscape Following Insufficient Radiofrequency Ablation in Hepatocellular Carcinoma: Through Multi-Omics and Experimental Validation

doi: 10.2147/JHC.S517528

Figure Lengend Snippet: ( A ) SNV mutations in two genes (EIF5B and ARF4) in the risk signature model. ( B ) The co-occurrence probabilities of the two key genes and the ten most mutated genes were analyzed. ( C ) Detection of gain/loss of CNV in two key genes. ( D ) Detection of gain/loss of CNV frequency in two key genes. ( E ) Correlations between key genes and different molecular features of hepatocellular carcinoma were analyzed. ( F ) Risk genes were analyzed for correlation with immune cells, EIF5B and ARF4 were significantly and positively correlated with the majority of immune cells. ( G ) The correlation between two key genes, EIF5B and ARF4. ( H ) Feature plot of EIF5B and ARF4 expression after recognition of cell clusters by umap. ( I ) Violin plots showing the distribution of EIF5B and ARF4 in different immune cells. ( J ) The pathways in which the two key genes were mainly enriched were analyzed by GSEA.

Article Snippet: Next, tissue sections were incubated overnight at 4°C with anti-ARF4 primary antibody (A7644, Abclonal) and EIF5B (A15123, Abclonal) at a dilution of 1:200.

Techniques: Expressing

Journal: Journal of Hepatocellular Carcinoma

Article Title: Decoding ARF4 and EIF5B-Based Prognostic Signatures and Immune Landscape Following Insufficient Radiofrequency Ablation in Hepatocellular Carcinoma: Through Multi-Omics and Experimental Validation

doi: 10.2147/JHC.S517528

Figure Lengend Snippet: ( A ) Variations in mRNA expression levels of ARF4 and EIF5B in HepG2 and SNU449 cell lines, as well as mouse tissues, under control conditions and following heat treatment. ( B ) Analysis of ARF4 and EIF5B at the protein levels in HepG2 and SNU449 cell lines. ( C ) Quantification of ARF4 and EIF5B protein levels in mouse tissues before and after exposure to IRFA. ( D ) Schematic representation of the IRFA model used for constructing subcutaneous tumors in mice. ( E ) Histopathological analysis and immunohistochemical staining of ARF4 and EIF5B in HCC sections from normal tumors and tumors post-IRFA in a murine model. ( F ) Fluorescence microscopy images depicting ARF4 and EIF5B expression patterns in HepG2, SNU449 cell lines and in mouse tissues. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Next, tissue sections were incubated overnight at 4°C with anti-ARF4 primary antibody (A7644, Abclonal) and EIF5B (A15123, Abclonal) at a dilution of 1:200.

Techniques: Expressing, Control, Immunohistochemical staining, Staining, Fluorescence, Microscopy

Journal: Journal of Hepatocellular Carcinoma

Article Title: Decoding ARF4 and EIF5B-Based Prognostic Signatures and Immune Landscape Following Insufficient Radiofrequency Ablation in Hepatocellular Carcinoma: Through Multi-Omics and Experimental Validation

doi: 10.2147/JHC.S517528

Figure Lengend Snippet: Assessment of the impact of ARF4 and EIF5B knockdown on the invasive, migratory, and proliferative capacities of HepG2 and SNU449 cells. ( A and E ) RT-qPCR was employed to quantify the efficacy of ARF4 and EIF5B knockdown in HepG2 and SNU449 cells. ( B and F ) The CCK-8 assay was conducted to evaluate the proliferation of HepG2 and SNU449 cells. ( C and G ) Cell migration was assessed via transwell assays. ( D and H ) Cell invasion was assessed via transwell assays. ( I and J ) The colony-forming assay was employed to assess the proliferative potential of HepG2 and SNU449 cells. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Next, tissue sections were incubated overnight at 4°C with anti-ARF4 primary antibody (A7644, Abclonal) and EIF5B (A15123, Abclonal) at a dilution of 1:200.

Techniques: Knockdown, Quantitative RT-PCR, CCK-8 Assay, Migration

Journal: Journal of Hepatocellular Carcinoma

Article Title: Decoding ARF4 and EIF5B-Based Prognostic Signatures and Immune Landscape Following Insufficient Radiofrequency Ablation in Hepatocellular Carcinoma: Through Multi-Omics and Experimental Validation

doi: 10.2147/JHC.S517528

Figure Lengend Snippet: ( A ) Immunofluorescence staining delineated alterations in the expression profiles of EMT-related genes, especially highlighting the dynamics of E-cadherin, Snail, and vimentin (depicted in green) within HCC cells. The cellular nuclei were concurrently counterstained with DAPI (blue). ( B ) Heatmap of gene changes pre and post ablation analysis using transcriptome sequencing. ( C ) Analysis of transcriptome sequencing reveals volcanic picture in gene expression pre- and post-ablation. ( D ) Transcriptome sequencing was used to analyze the changes in the differentially expressed genes EIF5B and ARFs family. ( E and F ) GO and KEGG pathway enrichment analysis. ( G ) GSEA of ARF4 and EIF5B-associated signaling pathways. ( H ) Shared pathways between ARF4 and EIF5B. ( I ) Pathway enrichment heatmap (NES) between ARF4 and EIF5B.

Article Snippet: Next, tissue sections were incubated overnight at 4°C with anti-ARF4 primary antibody (A7644, Abclonal) and EIF5B (A15123, Abclonal) at a dilution of 1:200.

Techniques: Immunofluorescence, Staining, Expressing, Sequencing, Gene Expression, Protein-Protein interactions