Journal: Alzheimer's & Dementia

Article Title: ZBP1‐mediated Treg cell dysfunction exacerbates neuroimmune dysregulation in Alzheimer's disease under metabolic stress

doi: 10.1002/alz.71896

Figure Lengend Snippet: ZBP1 impairs Treg‐mediated suppression of microglial activation, phagocytic dysfunction, and astrocytic C3 activation under metabolic stress. A, Schematics of the transwell co‐culture systems used to assess the effects of Tregs on primary microglia and astrocytes. B, C, Representative immunoblots and densitometric quantification of iNOS and MRC1 in primary microglia in each group ( n = 4 per group). D, Representative IF images of primary microglia stained for IBA1 (green) and iNOS (red) under the indicated conditions. Nuclei are counterstained with DAPI. Scale bar, 50 µm. E, F, Quantification of the percentage of iNOS + primary microglia and iNOS fluorescence intensity within IBA1 + microglia ( n = 6 per group). G, Schematic of the latex bead phagocytosis assay in primary microglia stimulated with LPS plus IFN‐γ and co‐cultured with Tregs. H, Representative flow‐cytometric plots of fluorescent latex bead uptake by primary microglia after LPS plus IFN‐γ stimulation in the presence or absence of WT or ZBP1‐KO Treg cells, with or without HGHP exposure. I, Quantification of the percentage of phagocytic microglia in the indicated groups ( n = 5 per group). J, Representative IF images of primary astrocytes stained for GFAP (green) and C3 (purple) under the indicated conditions. Nuclei were counterstained with DAPI. Scale bar, 50 µm. K, L, Quantification of the percentage of C3+ astrocytes and C3 fluorescence intensity within GFAP+ astrocytes ( n = 6 per group). Data are presented as mean ± standard deviation; NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001. Aβ, amyloid beta; DAPI, 4′,6‐diamidino‐2‐phenylindole; GFAP, glial fibrillary acidic protein; HGHP, high glucose and high palmitate; IBA1, ionized calcium‐binding adapter molecule 1; IF, immunofluorescence; IFN‐γ; interferon gamma; iNOS, inducible nitric oxide synthase; KO, knockout; LPS, lipopolysaccharide; MRC1, mannose receptor 1; Treg, regulatory T cell; WT, wild type; ZBP1, Z‐DNA‐binding protein 1.

Article Snippet: To evaluate microglial phagocytic activity, primary microglia in the lower chamber were stimulated with interferon gamma (IFN‐γ; 20 ng/mL; I4777, Sigma‐Aldrich) and lipopolysaccharide (LPS; 100 ng/mL; HY‐D1056, MedChemExpress) for 24 hours under Transwell co‐culture conditions.

Techniques: Activation Assay, Co-culture Assay, Western Blot, Staining, Fluorescence, Phagocytosis Assay, Cell Culture, Standard Deviation, Binding Assay, Immunofluorescence, Knock-Out

Journal: Respiratory Research

Article Title: The Tie-2 agonist PMC-403 inhibits endothelial damage induced by e-cigarette vapor or LPS

doi: 10.1186/s12931-026-03928-4

Figure Lengend Snippet: A HUVECs were stimulated with ECVE nic or control (EBM-2) for 12 hours (incubation). Then, 100 ng/ml LPS with co-factors LBP and CD14 or co-factors alone (sham) were added and incubated for another 18 hours. Created in BioRender. Stahl, K. (2026) https://BioRender.com/nu4iwxz . B Angpt-2 concentrations in the supernatant were measured by ELISA when LPS was added as well as 12 and 18 hours after LPS or sham stimulation and are shown over time. Each group consisted of n =6. Mixed ANOVA with according post-tests were performed. C 18 hours after stimulation with LPS or sham Angpt-2 mRNA transcription was measured by qPCR. Each group consisted of n =9. D Additionally, the Angpt-2/Tie2 mRNA ratio is shown after initial ECVE nic incubation and 18 hours of LPS or sham stimulation. ECVE nic / sham consisted of n=8, all other groups consisted of n =9. Groups were analyzed by Kruskal-Wallis test with post-hoc Dunn test. E TER was measured at different time points by an ECIS system. HUVECs were pre-incubated with ECVE nic ( n =7) or control ( n =8) for 12 hours. Then half of the medium was changed and cells were stimulated with LPS. Results are shown as dot plots with mean ± SD. A linear mixed-effects model was used for analysis. P -values < 0.05 are considered significant. Abbreviations: Angpt-2 - Angiopoietin-2, ECIS - Electric cell-substrate impedance sensing, ECVE – e-cigarette vapor extract, HUVECs – human umbilical vein endothelial cells, LPS - lipopolysaccharide, SD – standard deviation, TER – transendothelial electrical resistance

Article Snippet: Unless specified otherwise, cells were stimulated with 100 ng/ml lipopolysaccharide (LPS) from E. coli serotype O111:B4 (L4130, Sigma-Aldrich, St. Louis, MO) or vehicle (EBM-2), 100 ng/ml lipopolysaccharide binding protein (LBP) (870-LP-025, R&D Systems, Minneapolis, MN), 100 ng/ml CD14 (SRP3149, Sigma–Aldrich, St. Louis, MO), 500 ng/ml Angpt-1 (923-AN/CF, R&D Systems, Minneapolis, MN), electronic cigarette vapor extract (ECVE, see below) and/or different concentrations (e.g. 0.3, 1, 3–10 μg/ml) of the Tie2 agonist PMC-403 (PharmAbcine, Daejeon, Republic of Korea).

Techniques: Control, Incubation, Enzyme-linked Immunosorbent Assay, Analysis, Electric Cell-substrate Impedance Sensing, Standard Deviation