Journal: Redox Biology

Article Title: GLP-1R activation restores Gas6-driven efferocytosis in senescent foamy macrophages to promote neural repair

doi: 10.1016/j.redox.2025.103857

Figure Lengend Snippet: AMPK phosphorylation is required for Ex-4-induced activation of the Gas6/Axl pathway A, WB for p-AMPK and AMPK in BMDMs treated with MD, Ex-4 and Compound 5D. B, Densitometry analysis of p-AMPK. C, Molecular docking model of GLP-1R and AMPK. D , Co- IP for GLP-1R and AMPK. E-G , RMSD, Rg, and RMSF analyses of the GLP-1R–AMPK complex. H and I, Hydrogen bond counts and SASA of the complex. J and K, MM/GBSA energy contribution of key residues. L, FEL analysis of the complex. M, Chemical structure of BML-275. N, WB of Gas6 and p-Axl after BML-275 treatment. O and P, Densitometric quantification of Gas6 and p-Axl. Q, IF images of p-Axl (green) and Gas6 (pink) in BMDMs; scale bar = 100 μm. R , Quantification of fluorescence intensity. S, IF of IL-6 (green) and p21 (red) in BMDMs; scale bar = 100 μm. T, Quantification of IL-6 and p21. ∗, p < 0.05, ∗∗, p < 0.01, and ∗∗∗, p < 0.001.

Article Snippet: For functional assays, BMDMs were pretreated with either the p53-specific inhibitor Pifithrin-β (PFT-β, 10 μM for 24 h; HY-16702A, MedChemExpress), Exendin-4 (Ex-4, 100, 200, or 400 nM for 24 h; HY-13443, MedChemExpress), compound 5d (1 μM for 24 h, HY-101116, MedChemExpress), recombinant Mouse Gas6 (rGas6, 500 ng/mL for 2h; 986-GS-025/CF, R&D Systems, Minneapolis, MN, USA) or BML-275 (1 μM for 24 h; HY-13418A, MedChemExpress), followed by stimulation with MD (1 mg/mL) for 12 h.

Techniques: Phospho-proteomics, Activation Assay, Co-Immunoprecipitation Assay, Fluorescence

Journal: International journal of molecular sciences

Article Title: Stress-Induced Cholesterol Metabolic Dysregulation and Differentiation Trajectory Shift in Oligodendrocytes Synergistically Drive Demyelination.

doi: 10.3390/ijms26083517

Figure Lengend Snippet: Figure 3. AMPK pathway mediates abnormal cholesterol metabolism in OLGs. (A) Fold change in cholesterol metabolism genes in OPCs and OLGs under stress. Y-axis shows gene log2fc values. Cholesterol genes in cell were selected based on a p < 0.05. (B) KEGG pathway enrichment for upregulated DEGs in OLGs after 14 days of stress, focusing on environmental-information-processing pathways. (C) Western blot images and densitometric quantification of Ampkα2, p-Ampkα2, and Mbp in the hypothalamus. n = 4, values are mean ± SEM, ** p < 0.01, *** p < 0.001 vs. control. (D) Co-IF staining of OLGs (OLIG2+) and p-Ampkα2 in the hypothalamus (n = 6). Stress increases p-Ampkα2 expression around OLG nuclei at different time points, Bar = 27.6 µm. (E) Microinjection schematic into the lateral ventricle of mice (top: injection diagram, bottom: hematoxylin and eosin (HE) staining). Bar = 800 µm. (F) Western blot images and quantification of Ampkα2, p-Ampkα2, and Mbp levels (n = 6) in the hypothalamus, with cholesterol levels (n = 4) measured. The RS_14d+BML- 275 group showed lower p-Ampkα2 and Mbp levels, and increased cholesterol compared to the control. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The AMPK inhibitor BML-275 (dorsomorphin dihydrochloride, Cat# HY-13418, MedChemExpress, Monmouth Junction, NJ, USA) was delivered via intracerebroventricular stereotaxic injection (due to the large dosage, direct injection into hypothalamic tissue will excessively damage normal tissue and affect its function).

Techniques: Western Blot, Control, Staining, Expressing, Microinjection, Injection

Journal: International journal of molecular sciences

Article Title: Stress-Induced Cholesterol Metabolic Dysregulation and Differentiation Trajectory Shift in Oligodendrocytes Synergistically Drive Demyelination.

doi: 10.3390/ijms26083517

Figure Lengend Snippet: Figure 4. Stress induced abnormal cholesterol transport and differentiation trajectory in OLGs. (A) Co-IF staining of OLGs (OLIG2+) and cholesterol (BODIPY 480/508) in the hypothalamus (n = 6). The RS_14d+BML-275 group shows increased cholesterol accumulation around OLG nuclei, Bar = 27.6 µm. (B) Differential fold changes in myelin-related genes in OLGs at different stress timepoints. Selection for myelin genes in cell is based on DEGs with p < 0.05. (C) Dot plot of cholesterol transport gene expression to the ER in OLGs after 14 days of stress, showing reduced expression of cholesterol transport genes (Lrp6, Srebf2, Npc2, Sec23a, Scaper, Apoe, Lpcat3). (D) Differentiation trajectory of OLGs in the 3-day stress group (top) and cell monocle_state tree structure trajectory (bottom). The differentiation trajectory arranges cells based on their distance from the default starting point of the pseudotime axis using Monocle2. (E) Tree-structured trajectory distribution of OLG subpopulations in the 3-day stress group. (F) Differentiation trajectory of OLGs in the 14-day stress group (top) and the cell monocle_state tree structure trajectory (bottom). (G) OLG subpopulation tree structure trajectory in the 14-day stress group.

Article Snippet: The AMPK inhibitor BML-275 (dorsomorphin dihydrochloride, Cat# HY-13418, MedChemExpress, Monmouth Junction, NJ, USA) was delivered via intracerebroventricular stereotaxic injection (due to the large dosage, direct injection into hypothalamic tissue will excessively damage normal tissue and affect its function).

Techniques: Staining, Selection, Gene Expression, Expressing