Journal: Bioactive Materials

Article Title: Extracellular vesicles derived from salivary gland stem cells cultured on microwell scaffolds loaded with WNT3A promote the recovery of salivary gland function damaged by radiation via the YWHAZ-PI3K-AKT pathway

doi: 10.1016/j.bioactmat.2025.06.024

Figure Lengend Snippet: Functional validation of YWHAZ as a key effector in 3D WNT -EV-mediated restoration of irradiated mouse SGOs. (A) Representative bright-field, H&E, and PAS images of SGOs under five conditions: untreated (Control), irradiated + PBS (PBS), irradiated + 3D WNT -EVs derived from sgEpSCs transfected with YWHAZ siRNA (YWHAZ siRNA-EVs), irradiated + 3D WNT -EVs derived from sgEpSCs transfected with scrambled siRNA (scrambled siRNA-EVs), and irradiated + 3D WNT -EVs (3D WNT -EVs). SGOs treated with PBS or YWHAZ siRNA-EVs exhibited smaller size and cystic morphology, while those treated with scrambled siRNA-EVs or 3D WNT -EVs showed regenerative end-bud structures. H&E staining showed keratin pearl-like structures in the PBS group, while PAS staining revealed mucin recovery in all EV-treated groups, with greater intensity in scrambled and 3D WNT -EV conditions. Scale bars, 100 μm. (B) Representative immunofluorescence images of phosphorylated AKT (p-AKT) in SGOs under each treatment condition. SGOs treated with 3D WNT -EVs or scrambled siRNA-EVs showed elevated p-AKT levels compared to the PBS and YWHAZ siRNA-EVs groups. DAPI was used for nuclear counterstaining. Scale bars, 100 μm. (C) Cell viability, size of mouse SGOs and mucin area in EV-treated groups compared with the control and PBS groups. (D) Quantification of p-AKT + in SGOs shown in (B). Individual data points represent biological or technical replicates, per group ranging from 3 to 9. Error bars indicate SD. Statistical significance was determined using one-way ANOVA followed by Tukey's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: RNA interference (RNAi) was performed in human salivary gland–derived epithelial stem cells (sgEpSCs) using a Silencer® Pre-designed siRNA targeting human YWHAZ (ID: 107095, Ref: AM16708, Lot: ASO2NMXZ) and a non-targeting negative control siRNA (Ref: AM4611, Lot: ASO2NAC8), both purchased from Invitrogen (Thermo Fisher Scientific, USA).

Techniques: Functional Assay, Biomarker Discovery, Irradiation, Control, Derivative Assay, Transfection, Staining, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: IFN-γ-mediated suppression of ANGPT2-Tie2 in endothelial cells facilitates tumor vascular normalization during immunotherapy

doi: 10.3389/fimmu.2025.1551322

Figure Lengend Snippet: IFN-γ-mediated activation of STAT1 is required for the reduction in Tie2-ANGPT2 expression. (A–C) HUVECs and HPMECs were pretreated with a JAKi (ruxolitinib) for 24 h before being stimulated with IFN-γ (1000 IU/mL) for an additional 24 hours. mRNA levels of TEK (A) and ANGPT2 (B) were assessed by RT-qPCR. (C) Protein expression levels of Tie2, ANGPT2, and JAK1 were assessed using immunoblotting techniques. (D–F) HUVECs and HPMECs were transfected with siSTAT1 or siNC and subsequently stimulated with IFN-γ for 24 hours. mRNA expression levels of TEK (D) and ANGPT2 (E) were measured via RT-qPCR. The p values were analyzed via one-way ANOVA. (F) Protein levels of Tie2, ANGPT2, phosphorylated STAT1 (Y701), and total STAT1 were assessed using immunoblotting techniques. (G, H) The putative binding sites of the STAT1 binding motif in the −2 kb TEK promoter (G) or ANGPT2 promoter (H) were predicted via the JASPAR database. ChIP-qPCR confirmed the binding sites of STAT1 in the promoters of the TEK and ANGPT2 genes.

Article Snippet: siRNAs designed to target STAT1 were bought from RiboBio (Guangzhou, China).

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Western Blot, Transfection, Binding Assay, ChIP-qPCR

Journal: Frontiers in Immunology

Article Title: IFN-γ-mediated suppression of ANGPT2-Tie2 in endothelial cells facilitates tumor vascular normalization during immunotherapy

doi: 10.3389/fimmu.2025.1551322

Figure Lengend Snippet: IFN-γ inhibits the angiogenic properties of HUVECs. (A) The proliferation of HUVECs and HPMECs treated with IFN-γ (1000 IU/mL) was assessed employing a CCK-8 assay at 24, 48, and 72 hours post-culture. (B) Migration assays were conducted on HUVECs and HPMECs after stimulation with IFN-γ (1000 IU/mL) for 24 h. Scale bar: 50 μm. (C–F) Tube formation assays of HUVECs and HPMECs. Primary HUVECs were seeded in ECM and matrixgel. Photographs taken 10 h post-seeding were analyzed. HUVECs exhibited a tubular morphology after treatment with IFN-γ (1000 IU/mL) or F-ara-A (fludarabine, a STAT1 inhibitor) alone (C) or LY294002 alone (E) or in combination. Scale bar: 400 μm. (D, F) Quantification of tube formation, encompassing total length, number of branches, number of junctions, and number of meshes, is presented. The p values were determined via one-way ANOVA.

Article Snippet: siRNAs designed to target STAT1 were bought from RiboBio (Guangzhou, China).

Techniques: CCK-8 Assay, Migration

Journal: Frontiers in Immunology

Article Title: IFN-γ-mediated suppression of ANGPT2-Tie2 in endothelial cells facilitates tumor vascular normalization during immunotherapy

doi: 10.3389/fimmu.2025.1551322

Figure Lengend Snippet: IFN-γ-mediated suppression of ANGPT2-Tie2 in endothelial cells facilitates tumor vascular normalization during immunotherapy. Diagrammatic representation of the mechanism by which PD-L1 blockade therapy modulates IFN-γ to influence the tumor vasculature in ECs through the ANGPT2-Tie2 signaling pathway. IFN-γ motivates the JAK1/2-STAT1 signaling pathway, thereby inhibiting the expression of ANGPT2 and Tie2. Following this process, phosphorylated STAT1 binds to the promoter regions of the TEK and ANGPT2 genes, thereby suppressing their gene expression. Furthermore, IFN-γ phosphorylates FOXO1 through activating the PI3K-AKT pathway, thereby reducing the nuclear activation of FOXO1 on the ANGPT2 gene. Consequently, the synthesis of the Tie2 and ANGPT2 proteins is diminished, contributing to the suppression of the ANGPT2-Tie2 signaling pathway. This effect subsequently suppresses the migration, proliferation, and tube formation of ECs, ultimately contributing to the suppression of tumor angiogenesis and vascular destabilization.

Article Snippet: siRNAs designed to target STAT1 were bought from RiboBio (Guangzhou, China).

Techniques: Expressing, Gene Expression, Activation Assay, Migration