Journal: Molecular Therapy. Nucleic Acids

Article Title: LOXL1 exerts oncogenesis and stimulates angiogenesis through the LOXL1-FBLN5/αvβ3 integrin/FAK-MAPK axis in ICC

doi: 10.1016/j.omtn.2021.01.001

Figure Lengend Snippet: RGD domain in FBLN5 is indispensable to the proangiogenic function of LOXL1 in ICC by binding to the αvβ3 integrin (A) pUC57, pFBLN5 RGD , and pFBLN5 RGE were transfected into RBE cells, and the efficiency of transfection was measured by western blot. A higher level of FBLN5 protein was contained in the supernatant of pFBLN5 transfected groups compared to pUC57 groups. (B and C) The roles of RGD domain and the αvβ3 integrin in the proangiogenic process of GST-LOXL1 CD were detected through tube-formation assays. Cyclo (-RGDfK) (50 ng/μL) was used to block the αvβ3 integrin. Scale bars, 100 μm (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). (D) Cell lysates of VECs were collected for testing phosphorylation levels of FAK and Erk1/2 by western blot. (E) Colocalization of LOXL1, FBLN5, and the αvβ3 integrin along with the vessel-like structure in the extracellular matrix of ICC tumor tissues. Scale bars, 100 μm.

Article Snippet: The inhibitor of the αvβ3 integrin, cyclo (-RGDfK), was purchased from MedChemExpress (MCE; USA) and diluted to 50 μg/μL for stock.

Techniques: Binding Assay, Transfection, Western Blot, Blocking Assay, Phospho-proteomics

Journal: Nature Communications

Article Title: Metabolism archetype cancer cells induce protumor TREM2 + macrophages via oxLDL-mediated metabolic interplay in hepatocellular carcinoma

doi: 10.1038/s41467-025-62132-y

Figure Lengend Snippet: A UMAP of the Scissor-selected cells (left). Barplot shows the distribution of Scissor + cells across different myeloid populations and conditions (right). B Forestplot shows the hazard ratios and 95% confidence intervals for different myeloid cluster signatures and clinical information according to a multivariable Cox model in the TCGA-LIHC cohort ( n = 353 patients). Squares represent the hazard ratios, and the horizontal bars extend from the lower limits to the upper limits of the 95% confidence intervals of the estimates of the hazard ratios. C Dotplot displays the DEGs between Scissor + cells and all other cells. D Dotplot displays the expression correlation of SPP1 with other genes in myeloid cells (left). Heatmap shows the expression correlation of indicated genes with SPP1 in myeloid cells from adjacent non-tumor (ANT) and tumor samples (right). E Representative images and quantification of TREM2, SPP1, and CD68 immunofluorescence staining on human HCC sections. Representative cells that denote the co-upregulation of TREM2 and SPP1 in CD68 + TAMs are circled by a dotted line. Scale bar, 50 μm. F Kaplan–Meier survival analysis of HCC patients from the TCGA-LIHC cohort categorized into groups based on normalized TREM2 and SPP1 expression. The cutpoints for patient grouping were calculated by the surv_cutpoint function from the survminer R package. G Barplot shows the mean expression of SPP1 across different myeloid cell populations (upper) and in TAMs from ICB-R and ICB-NR HCC samples (lower). H Dotplot shows the expression of SPP1 ligands in CD8 + T cells (above) or cancer cells (below) from pre-ICB, ICB-NR, and ICB-R HCC scRNA-seq samples. I Schematic of the interaction between TREM2 + TAMs and CD8 + T cells or cancer cells in immunotherapy-resistant HCC. J Heatmap shows the relative expression of indicated genes in isolated TREM2 + /TREM2 − TAMs. K Flow cytometry of IFNγ expression in CD8 + T cells co-cultured with human HCC-isolated TREM2 - TAMs or TREM2 + TAMs ± SPP1 antibody (1 μg/mL). CD8 + T cells were isolated from human PBMC and then activated with anti-CD3/CD28 + IL-2 (50 U/mL) before co-culture assays. L Violin plots display the expression of IFNG and TNF in CD8 + T cells from ICB-NR or ICB-R scRNA-seq samples, with two-tailed Wilcoxon-test statistics. M Dotplot shows the relative expression of IFNG and TNF in CD8 + T cell clusters. N Tissue preference of CD8 + T cell clusters, revealed by odds ratio (OR) value. O Relative cell viability (mean of n = 3 biological replicates) for Hep3B cells treated with a half-log dilution series of TNF (2.5–250 ng/mL) and IFNγ (1–100 ng/mL), cocultured with human HCC-isolated TREM2 - TAMs or TREM2 + TAMs ± SPP1 antibody (1 μg/mL). P Relative cell viability of Hep3B cells treated with mock, SPP1 (50 ng/mL), integrin inhibitor TFA (100 μM), TNF (250 ng/mL) plus IFNγ (100 ng/mL) (TNF/IFNγ), TNF/IFNγ plus SPP1, or TNF/IFNγ plus SPP1 plus TFA (100 μM) for 24 h. Q Relative cell viability for patient-derived HCC organoid with indicated treatment. Data represent the mean ± SD, n = 5 biological replicates in ( P ), n = 6 biological replicates in ( K , Q ). Statistical significance was determined by two-tailed Wald test ( B ), two-tailed unpaired t test ( K , P , Q ), two-tailed Wilcoxon signed rank test ( L ), and log rank test ( F ). Schematic in I was created in BioRender. Chu, T. (2025) https://BioRender.com/r2v4jgs . Source data are provided as a Source Data file.

Article Snippet: Then, TNF (100 ng/mL) + IFNγ (100 ng/mL) (TNF/IFNγ), TNF/IFNγ plus SPP1 (50 ng/mL) or a combination of TNF/IFNγ, SPP1, and integrin inhibitor TFA (HY-100445A, MCE, 50 μM) was added to the culture medium.

Techniques: Expressing, Immunofluorescence, Staining, Isolation, Flow Cytometry, Cell Culture, Co-Culture Assay, Two Tailed Test, Derivative Assay