Journal: Frontiers in Immunology
Article Title: Single-cell atlas of human skin implicates APOE pro-inflammatory signaling in diabetic foot ulcers
doi: 10.3389/fimmu.2025.1591944
Figure Lengend Snippet: APOE+ fibroblasts promote fibrosis and inflammation in DFU. (A) Representative images showing histological differences between human healthy donor (HD) and diabetic foot ulcer (DFU) skin tissues assessed by HE staining (left) and APOE expression visualized by IHC (right). Insets display magnifications of different regions in the skin. Scale bar in the original images: 500 μm, 2000 μm. Scale bar in the magnified images: 50 μm. Upper: Cellular infiltration was quantified in HE-stained sections by measuring the percentage of infiltrated area. Lower: APOE expression was quantified through positive area percentage analysis of IHC staining. (B) Western blot analysis and quantitative comparison of APOE protein in human HD and DFU skin tissues. (C) RT-qPCR analysis of mRNA levels of α-SMA, COL1A1 and COL3A1 in human HD and DFU skin tissues. (D) Western blot analysis and quantitative comparison of the NF-κB and JAK1/Stat3 signaling pathways in human HD and DFU skin tissues. (E) Western blot analysis and quantitative comparison of the NF-κB and JAK1/Stat3 signaling pathways in human fibroblasts treated with APOE3 at concentrations of 10 ng/mL or 25 ng/mL. (F) Western blot analysis and quantitative comparison of the NF-κB and JAK1/Stat3 signaling pathways in human fibroblasts treated with APOE3 at a concentration of 10 ng/mL for 6 h, 12 h or 24 h. Data are presented as mean ± SD from three independent biological replicates (n = 3). * p < 0.05, *** p < 0.001.
Article Snippet: Cells were treated with human recombinant APOE3 protein (MedChemExpress, China) at 10 ng/mL or 25 ng/mL for 24 hours, or at 10 ng/mL for 6, 12, or 24 hours.
Techniques: Staining, Expressing, Immunohistochemistry, Western Blot, Comparison, Quantitative RT-PCR, Protein-Protein interactions, Concentration Assay
