Journal: Bioactive Materials
Article Title: Pathology-inspired collagen-binding thermosensitive micelle drops enable prolonged and efficient treatment of fungal keratitis
doi: 10.1016/j.bioactmat.2025.04.011
Figure Lengend Snippet: Therapeutic mechanism of PX-TA-AmB in FK via MAPK6/PI3K/AKT pathway modulation. (A) Volcano plot of the differentially expressed genes. (B) KEGG pathway enrichment analysis of the differentially expressed genes. (C) Heatmap of the differentially expressed genes related to the inflammatory response, signaling pathway regulation, and corneal scarring before and after PX-TA-AmB treatment. (D, E) GSEA with the indicated KEGG and Reactome gene sets. (F, G, H) MAPK6, α-SMA, and LOX mRNA expression levels in treated versus control corneal tissues. (I) Representative Western blot images of p-AKT, AKT, p-PI3K, PI3K, MAPK6, Vinculin, and β-actin proteins in different treatment groups. (J) Representative Western blot images of IL-1β and MMP9 in different treatment groups. (K) Representative Western blot images for functional validation of the role of AKT pathway in PX-TA-AmB-mediated downregulation of IL-1β and MMP9 using the AKT activator SC79. (L) Immunofluorescence assessment of α-SMA, MMP9, CD206, CD86, and Ly6G under different treatments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Mean ± SD, n = 3; one-way ANOVA.
Article Snippet: The total protein was separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto a 0.45 μm pore size polyvinylidene fluoride (PVDF) membrane (Millipore, Germany); this membrane was then blocked with 5 % milk at room temperature and incubated with primary antibodies overnight at 4 °C; these antibodies included p-PI3K (Cell Signaling Technology, 4228, 1:1000), PI3K (Cell Signaling Technology, 4257, 1:1000), p-Akt (ABclonal, AP1453, 1:1000), Akt (ABclonal, A18675, 1:2000), MAPK6 (Abcam, ab53277, 1:1000), IL-1β (Abcam, ab9722, 1:1000), MMP9 (Proteintech, 10375-2-AP, 1:1000), Vinculin (Proteintech, 26520-1-AP, 1:10000) and β-Actin (ABclonal, AC026, 1:20000).
Techniques: Expressing, Control, Western Blot, Functional Assay, Biomarker Discovery, Immunofluorescence
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