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