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: iScience

Article Title: ZDHHC17-mediated IKKβ palmitoylation suppresses NF-κB signaling to mitigate the progression of liver fibrosis

doi: 10.1016/j.isci.2026.116360

Figure Lengend Snippet: ZDHHC17 deficiency stimulates pro-inflammatory cytokines production in macrophages leading to enhanced expression of fibro-genic genes in HSCs (A) THP-1 cells were induced to M0 macrophages by PMA and further polarized to M1 macrophages by treatment with LPS/IFN-γ. ZDHHC17 was over-expressed or knocked down in M1 macrophages by plasmid transfection or siRNA. Cells were collected 48 h post-transfection for detection. LX2 cells were seeded onto the lower chamber, and M1 macrophages were plated on the upper chamber of a 0.4 μm transwell membrane, which prevents direct cell contact, limiting cell interaction to secretory chemicals. (B and C) ZDHHC17 was significantly overexpressed in M1 macrophages by plasmid transfection (B) and decreased by small interfering RNAs (siRNAs) (C). Data are presented as the mean ± SD ( n = 6 independent experiments). mRNA expression of the pro-inflammatory cytokines (TNF-α, IL-1β, and IL-12β) were detected in ZDHHC17-overexpressed (D) or ZDHHC17-silenced (E) macrophages. Data are presented as the mean ± SD ( n = 6 independent experiments). ELISA detected serum TNF-α, IL-1β, and IL-12β in ZDHHC17-overexpressed (F) or ZDHHC17-silenced (G) macrophages. Data are presented as the mean ± SD ( n = 6 independent experiments). mRNA expression of α-SMA, COL1A1, TGF-β, and TIMP-1 was detected by qPCR in LX-2 cells following co-cultured with ZDHHC17-overexpressed (H) or silenced (I) macrophages. Data were presented as mean ± SD ( n = 6 independent experiments). ∗ p < 0.05 , ∗∗ p < 0.01 , ∗∗∗ p < 0.001 , ∗∗∗∗ p < 0.0001 by Student’s t test (B-I).

Article Snippet: LPS (Lipopolysaccharide) , MCE (Monmouth Junction, NJ, USA) , Cat# HY-D1056.

Techniques: Expressing, Plasmid Preparation, Transfection, Membrane, Enzyme-linked Immunosorbent Assay, Cell Culture