Journal: Journal of Extracellular Vesicles

Article Title: Extracellular Vesicle‐Mediated Nucleolin Transfer in Glioblastoma: A Targetable Axis Driving Blood‐Tumour Barrier Formation

doi: 10.1002/jev2.70268

Figure Lengend Snippet: Transfer of NCL from GBM‐derived EVs to brain capillary endothelial cells. (A) Schematic of the proteomic analysis for EVs derived from mouse normal astrocytes and U87MG cells. (B) PCA of the proteomic profile of astrocyte‐derived EVs and U87MG‐derived EVs. (C) Volcano plot showing proteins with differential abundance between astrocyte‐derived EVs and U87MG‐derived EVs. P < 0.001, |Log 2 FC| > 5. (D) Functional enrichment analysis of differentially expressed proteins carried by U87MG‐derived EVs. (E) Schematic of an in vitro BBB or BTB Transwell model, achieved by coculturing bEnd.3 cells with normal astrocytes, untreated U87MG cells, U87MG cells transfected with scrambled control shRNA (shCtrl) or NCL shRNA (shNCL), or U87MG cells preincubated with DMSO or 10 µM GW4869. After the coculture for 5 days, the surface expression of NCL on bEnd.3 cells were detected by Western blotting or flow cytometry. (F) Level of surface NCL on bEnd.3 cells after the indicated coculture. (G) The ratio of surface NCL‐expressing bEnd.3 cells after the indicated coculture. (H) EdU proliferation assay for bEnd.3 cells after the indicated coculture. (I) TUNEL assay for apoptotic bEnd.3 cells after the indicated coculture. (J) Migration of bEnd.3 cells after the indicated coculture. (K) Tube formation assay for bEnd.3 cells after the indicated coculture. (L) Schematic showing the incubation of bEnd.3 cells with 10 µg/mL EVs from normal astrocytes, untreated U87MG cells, or U87MG cells transfected with shCtrl or shNCL. After incubation for 12 h, surface expression of NCL on bEnd.3 cells were assessed using Western blotting or flow cytometry. (M) Level of surface NCL on bEnd.3 cells post‐incubation with EVs from the indicated cells. (N) The ratio of surface NCL‐expressing bEnd.3 cells post‐incubation with EVs from the indicated cells. (O) EdU proliferation assay for bEnd.3 cells after coculture with EVs from the indicated cells. (P) TUNEL assay for apoptotic bEnd.3 cells after coculture with EVs from the indicated cells. (Q) Migration of bEnd.3 cells after coculture with EVs from the indicated cells. (R) Tube formation assay for bEnd.3 cells after coculture with EVs from the indicated cells. (S) Schematic illustration of the establishment of an in vivo orthotopic GBM mouse model. Briefly, U87MG cells were engineered to express firefly luciferase and GFP (U87MG‐GFP‐Luc cells) and transfected with shCtrl or shNCL before being inoculated into the brains of BALB/c nude mice for 7 days. (T) IF staining of CD31 (red) and NCL (yellow) of brain sections from the orthotopic GBM mice. Nuclei were counterstained with DAPI (blue). N, non‐tumour region; T, tumour region (green). Scale bars, 50 µm. In vitro experiments were performed in triplicate and repeated three times. n = 3 for each group of in vivo studies. Graphs represented means ± SEM, and statistical significance was calculated by one‐way ANOVA (G, H, I, J, K, N, O, P, Q, and R). * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: After counterstaining with DAPI (Beyotime, CHN), confocal images were acquired using a confocal fluorescence microscope (Zeiss 980, DE).

Techniques: Derivative Assay, Functional Assay, In Vitro, Transfection, Control, shRNA, Expressing, Western Blot, Flow Cytometry, Proliferation Assay, TUNEL Assay, Migration, Tube Formation Assay, Incubation, In Vivo, Luciferase, Staining

Journal: Materials Today Bio

Article Title: Bioinspired lipid droplets nanoplatform for periodontitis therapy: Integrated antibacterial, mitochondrial repair, and immunomodulatory functions

doi: 10.1016/j.mtbio.2026.102808

Figure Lengend Snippet: Exploration of the anti-inflammatory and antioxidant mechanisms of GA@LDs-CRAMP in BMDMs . A) Immunofluorescence images of Nrf2 (red) and nuclei (blue) in LPS-stimulated BMDMs. Scale bar: 10 μm. B) Quantitative analysis of the Pearson colocalization coefficient between Nrf2 red fluorescence and DAPI-stained nuclear blue fluorescence (n = 5). C) Western blot validation and semiquantitative analysis of macrophage polarization markers (iNOS, Arg1) and inflammatory cytokines (IL-6). D) Western blot validation and semiquantitative analysis of relative expression of NF-κB pathway components (p65, p-p65, IκBα, p-IκBα) (n = 3). E) Western blot validation and semiquantitative analysis of relative expression of Nrf2 pathway components (Nrf2, Keap1, HO-1, NQO1) (n = 3). ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Synthesis conditions were screened by staining LDs with BODIPY 493/503 (5 μM, MedChemExpress, USA) and counterstaining nuclei with DAPI (7 μM, Beyotime, China).

Techniques: Immunofluorescence, Fluorescence, Staining, Western Blot, Biomarker Discovery, Expressing