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: Gas6 is a critical downstream target of Ex-4 in alleviating macrophage senescence and efferocytosis impairment A, SA-β-gal staining in BMDMs transfected with LV-shGas6; scale bar = 100 μm. B, Quantification of SA-β-gal-positive cells. C–G, Relative mRNA expression of SASP factors and senescence markers in LV-shGas6-transfected BMDMs. H, Live-cell imaging of BMDMs post-LV-shGas6 transfection; scale bar = 5 μm. I, IF images of NeuN (blue) and MAP2 (red) in neurons co-cultured with Gas6-deficient BMDMs; scale bar = 50/25 μm. J, Sholl analysis of neuronal branching. K, IF images of GFAP (green) and Aggrecan (red) in astrocytes co-cultured with Gas6-deficient BMDMs; scale bar = 100 μm ∗, 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: Staining, Transfection, Expressing, Live Cell Imaging, Cell Culture

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: Ex-4 attenuates macrophage senescence and efferocytosis dysfunction through activating Gas6 in SCI mice A, IF images of F4/80 (green), Gas6 (pink), F4/80 (blue), p-Axl (pink) at 7 dpi in spinal cords following AAV-shGas6 and Ex-4 treatment; scale bar = 200/100/50 μm. B, IF images of F4/80 (green), IL-6 (pink), F4/80 (blue), p21 (red); scale bar = 200/100/50 μm. C, IF images of F4/80 (green) and LAMP2 (pink); scale bar = 200/100/50 μm. D–H, Quantification of Gas6-, p-Axl-, IL-6-, p21-, and LAMP2-positive macrophage areas. ∗, 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:

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: Ex-4 suppresses glial scarring and promotes remyelination and axonal regeneration in SCI mice via the Gas6/Axl signaling pathway A, IF images of IBA-1 (pink) and GFAP (blue) at 7 and 28 dpi; scale bar = 100/50 μm. B, IF images of MBP (pink) and NF-200 (green); scale bar = 100/50 μm. C and D, Quantification of glial scar area at 7 dpi. E and F , Glial scar area at 28 dpi. G and H, Quantification of myelinated and axonal area at 7 dpi. I and J, Myelinated and axonal area at 28 dpi. ∗, 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:

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: Ex-4 promotes neuroprotection and functional recovery in SCI mice through the Gas6/Axl axis A, HE staining of spinal cords at 28 dpi; scale bar = 500/100 μm. B, Nissl staining at 28 dpi; scale bar = 500/100 μm. C, LFB staining at 28 dpi; scale bar = 500/100 μm. D-F, Quantification of lesion size, neuronal survival, and demyelination. G and H, Footprint analysis quantification at 28 dpi. I, Representative footprint images at 28 dpi. J, BMS scores across 28 dpi. ∗, 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: Functional Assay, Staining

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