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

Journal: Beilstein Journal of Nanotechnology

Article Title: Serum heat inactivation diminishes ApoE-mediated uptake of D-Lin-MC3-DMA lipid nanoparticles

doi: 10.3762/bjnano.16.57

Figure Lengend Snippet: Schematic representation of the heat treatment and ThT staining of ApoE. Binding of ThT to misfolded or aggregated proteins greatly enhances its fluorescence (A). The fluorescence of solutions containing ApoE3 or BSA following ThT staining as determined by spectrophotometry, after a 30 min incubation at room temperature (rt), 37, 56 and 75 °C ( n = 3) (B). Differences were considered statistically significant at p < 0.05 and were annotated as ns = non-significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 and **** = p ≤ 0.0001. The graphics in were generated with images provided by Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution 4.0 International License, https://creativecommons.org/licenses/by/4.0/

Article Snippet: Human recombinant ApoE3 was purchased from Fitzgerald Industries.

Techniques: Staining, Binding Assay, Fluorescence, Spectrophotometry, Incubation, Generated

Journal: Beilstein Journal of Nanotechnology

Article Title: Serum heat inactivation diminishes ApoE-mediated uptake of D-Lin-MC3-DMA lipid nanoparticles

doi: 10.3762/bjnano.16.57

Figure Lengend Snippet: Heat-mediated aggregation/denaturation prevents ApoE from binding to LNPs and reduces LNP uptake by cells. The fluorescence of HMEC-1 after uptake of LNP containing alexa fluor 647 labelled siRNA at different concentrations of recombinant ApoE3, BSA or medium supplemented with NHI FCS (A). The concentration of fluorescent siRNA was 1.725 pmol per well and uptake was measured after 4 h ( n = 3). The fluorescence of HMEC-1 cells after uptake of LNPs in medium supplemented with HI or NHI ApoE (B). Schematic representation of the bead capture and staining of ApoE bound LNPs (C). The fluorescence of ApoE3 bound to LNPs that are captured by magnetic beads after incubation in human serum, heat treated human serum and recombinant ApoE3 in PBS, as determined by flow cytometry (D) ( n = 2). Differences were considered statistically significant at p < 0.05 and were annotated as ns = non-significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 and **** = p ≤ 0.0001. The graphics in were generated with images provided by Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution 4.0 International License, https://creativecommons.org/licenses/by/4.0/

Article Snippet: Human recombinant ApoE3 was purchased from Fitzgerald Industries.

Techniques: Binding Assay, Fluorescence, Recombinant, Concentration Assay, Staining, Magnetic Beads, Incubation, Flow Cytometry, Generated

Journal: Beilstein Journal of Nanotechnology

Article Title: Serum heat inactivation diminishes ApoE-mediated uptake of D-Lin-MC3-DMA lipid nanoparticles

doi: 10.3762/bjnano.16.57

Figure Lengend Snippet: The uptake of MC3 LNPs is ApoE dependent and is affected by heat inactivation of FCS, while the uptake of C12 LNPs is not. Uptake of MC3 or C12 LNPs containing alexa fluor 647 labelled siRNA by HMEC-1, U87 and MDA-MB-231 in medium supplemented with NHI FCS, HI FCS or HI FCS with 1 µg/mL ApoE3. Analysed by flow cytometry ( n = 3). Differences were considered statistically significant at p < 0.05 and were annotated as ns = non-significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 and **** = p ≤ 0.0001.

Article Snippet: Human recombinant ApoE3 was purchased from Fitzgerald Industries.

Techniques: Flow Cytometry

Journal: Beilstein Journal of Nanotechnology

Article Title: Serum heat inactivation diminishes ApoE-mediated uptake of D-Lin-MC3-DMA lipid nanoparticles

doi: 10.3762/bjnano.16.57

Figure Lengend Snippet: Heat inactivation of FCS and endogenous ApoE diminishes the firefly luciferase knockdown efficacy of MC3 LNPs, but not of C12 LNPs. Firefly luciferase activity in U87 and MDA-MB-231 48 h after uptake of siLuc loaded MC3 or C12 LNPs in medium supplemented with NHI FCS, HI FCS or HI FCS with 1 µg/mL ApoE3. Luciferase activity was analysed by measuring bioluminescence ( n = 3). Differences were considered statistically significant at p < 0.05 and were annotated as ns = non-significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 and **** = p ≤ 0.0001.

Article Snippet: Human recombinant ApoE3 was purchased from Fitzgerald Industries.

Techniques: Luciferase, Knockdown, Activity Assay