α v β 5 integrin  (MedChemExpress)

Journal: Neurotherapeutics

Article Title: Cellular fibronectin exacerbates α-synuclein aggregation via integrin alpha4beta1 mediated PARP1 and SCD elevation

doi: 10.1016/j.neurot.2025.e00729

Figure Lengend Snippet: cFn induced PARP1 activation via EIIIA-integrin α4β1 interaction, which subsequently contributed to pathological α-syn aggregation and mitochondrial dysfunction in SH-SY5Y cells. (a) Representative western blots and quantification of nNOS and PARP1 in SH-SY5Y cells coated with cFn and treated with different receptor inhibitors (five independent replicates). (b) Quantification of NO release in SH-SY5Y cells treated with cFn or different receptor inhibitors (five independent replicates). (c) Representative western blots and quantification of the TX-insoluble α-syn oligomer and Ser129-phosphorylated α-syn in SH-SY5Y cells (five independent replicates). (d–e) Representative images and quantification of PARP1 (d) and γ-H2AX (e) staining in SH-SY5Y cells 12 ​h after L-NAME or PBS pretreatment (five independent replicates). DAPI (blue) was used for nuclear staining. Scale bar: 100 ​μm. (f) Representative western blots and quantification of PAR and PARP1 in SH-SY5Y cells coated with cFn or AF38Pep at different concentration (five independent replicates). (g) Representative coimmunoprecipitation images showing an interaction between Fn and the integrin α4 subunit in SH-SY5Y cells, and the quantification of Fn and α4 subunit level in the coimmunoprecipitation samples was performed (five independent replicates). All data are presented as the mean ​± ​s.e.m. Differences among 2 groups were analysed by Unpaired Student's t -test, while differences among multiple groups were analysed by ANOVA followed by Tukey's post hoc test. ∗P ​< ​0.05, ∗∗P ​< ​0.01, ∗∗∗P ​< ​0.0005, ∗∗∗∗P ​< ​0.0001, and ns, not significant.

Article Snippet: To prevent interactions between Fn and different receptors, SH-SY5Y cells were pretreated with the integrin α5β1 receptor inhibitor ATN-161 trifluoroacetate salt (MCE HY-13535A), the integrin α4β1 receptor inhibitor TR-14035 (MCE HY-15770), or the TLR4 inhibitor TLR4-IN-C34 (MCE HY-107575) following the manufacturers' protocols.

Techniques: Activation Assay, Western Blot, Staining, Concentration Assay

Journal: Neurotherapeutics

Article Title: Cellular fibronectin exacerbates α-synuclein aggregation via integrin alpha4beta1 mediated PARP1 and SCD elevation

doi: 10.1016/j.neurot.2025.e00729

Figure Lengend Snippet: cFn induced FAs and TAGs elevation by increasing SCD, which contributed to pathological α-syn aggregation. LC‒MS/MS Analysis generated data were performed in A-D. (a–b) Changes in FAs in the SNpc of cFn-injected mice compared with those in healthy mice were determined by hierarchical clustering analysis (a) and matchstick analysis (b) (n ​= ​6), (c–d) Changes in TAGs in the SNpc of cFn-injected mice compared with healthy mice were determined by hierarchical clustering analysis (c) and matchstick analysis (d) (n ​= ​6). In the image of hierarchical clustering analysis, red plots represent upregulation, and blue plots represent downregulation. In the matchstick analysis, red lines represent upregulation, while blue lines represent downregulation. ∗P ​< ​0.05, ∗∗P ​< ​0.01. FAs species was described in ; (e) Representative Oil Red O staining images and relative quantification of the positive area of lipid droplets in SH-SY5Y cells coated with cFn or treated with an integrin α4β1 inhibitor (five independent replicates). Oli Red O staining was detected in bright field channel. Scale bar: 25 ​μm. (f–g) The levels of TAGs (f) and FAs (g) in SH-SY5Y cells coated with cFn or treated with an integrin α4β1 inhibitor were measured (three independent replicates). (h) Representative immunoblots and quantification of SCD in the SNpc of the control, MPTP, MPTP ​+ ​AAV-Fn1-shRNA, and MPTP ​+ ​AAV-NC groups (n ​= ​5). (i) Representative immunoblots and quantification of SCD in SH-SY5Y cells coated with cFn or treated with an integrin α4β1 inhibitor (five independent replicates). (j) Representative western blots and quantification of SCD in the SNpc of mice injected with cFn or treated with AF38Pep (five independent replicates). (k–l) Representative coimmunoprecipitation images showing an interaction between SCD and the α4 subunit in the SNpc of healthy mice or cFn injected mice, and the quantification of α4 subunit or SCD level in the coimmunoprecipitation samples was performed (n ​= ​5). (m) Representative western blots and quantification of the TX-insoluble α-syn oligomer and Ser129-phosphorylated α-syn in α-syn-overexpressing SH-SY5Y cells coated with cFn or treated with an SCD inhibitor (five independent replicates). All data are presented as the mean ​± ​s.e.m. Differences among 2 groups were analysed by Unpaired Student's t -test, while differences among multiple groups were analysed by ANOVA followed by Tukey's post hoc test. ∗P ​< ​0.05, ∗∗P ​< ​0.01, ∗∗∗P ​< ​0.0005, ∗∗∗∗P ​< ​0.0001, and ns, not significant.

Article Snippet: To prevent interactions between Fn and different receptors, SH-SY5Y cells were pretreated with the integrin α5β1 receptor inhibitor ATN-161 trifluoroacetate salt (MCE HY-13535A), the integrin α4β1 receptor inhibitor TR-14035 (MCE HY-15770), or the TLR4 inhibitor TLR4-IN-C34 (MCE HY-107575) following the manufacturers' protocols.

Techniques: Generated, Injection, Staining, Quantitative Proteomics, Western Blot, Control, shRNA

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