Journal: bioRxiv

Article Title: APOE regulates the transport of GM1

doi: 10.1101/2024.04.02.587789

Figure Lengend Snippet: (a) Schematic illustration of determining the binding affinity between lipid structure and APOE using MST. (b) The binding affinity Kd between APOE3 and lipid structures with different compositions. (n>=4) (c) The binding affinity between APOE3 and lipid structures with different GM1 concentrations. (n>=3) (d) The binding affinity between APOE4 and lipid structures with different compositions. (n>=5) (e) The binding affinity between APOE4 and lipid structures with different GM1 concentrations. (n>=5) P-value: ns (0.05 < p <= 1), * (0.01 < p <= 0.05, ** (0.001 < p <= 0.01, *** (0.0001 < p <= 0.001, **** (p <= 0.0001). (a) is created with BioRender.com.

Article Snippet: To determine the binding affinity between APOE-lipoprotein and its receptor LDLR (MedChemExpress), we first labeled APOE3 and APOE4 with RED-NHS 2nd generation, as described earlier.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: APOE regulates the transport of GM1

doi: 10.1101/2024.04.02.587789

Figure Lengend Snippet: (a) The expression of LDLR on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n=4). (b) The expression of VLDLR on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n=4). (c) The expression of LRP1 on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n=3). (d) The expression of ApoER2 on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n=3). (e) Schematic illustration of determining the changes of APOE secondary structures. (f) The CD results of APOE 3 under the effect of different GM1 content on the lipid structures (n>=3). (g) The CD results of APOE 4 under the effect of different GM1 content on the lipid structures (n=3). (h) Schematic illustration of determining the binding affinity of APOE-enriched lipoprotein and APOE receptor LDLR using MST. (i) The binding affinity between APOE3 and lipid structures with different compositions. (n>=4) (j) The binding affinity between APOE4 and lipid structures with different compositions. (n>=3) P-value: ns (0.05 < p <= 1), * (0.01 < p <= 0.05, ** (0.001 < p <= 0.01, *** (0.0001 < p <= 0.001, **** (p <= 0.0001). (e) and (h) are created with BioRender.com.

Article Snippet: To determine the binding affinity between APOE-lipoprotein and its receptor LDLR (MedChemExpress), we first labeled APOE3 and APOE4 with RED-NHS 2nd generation, as described earlier.

Techniques: Expressing, Binding Assay

Journal: Journal of Molecular Endocrinology

Article Title: Upregulation of hepatic LRP1 by rosiglitazone: a possible novel mechanism of the beneficial effect of thiazolidinediones on atherogenic dyslipidemia

doi: 10.1530/jme-12-0119

Figure Lengend Snippet: Figure 3 The effect of rosiglitazone on ApoE uptake in HepG2 cells. HepG2 cells were treated with indicated concentrations of rosiglitazone for 48 h. Human recombinant ApoE3 was added to culture media and cells were incubated for 1 h. ApoE3 was reconstituted with lipid using DMPC before the treatment on HepG2 cells. Three independent experiments were performed for the representative figures. (A) Western blot analysis of ApoE3 in HepG2 cells incubated with or without added ApoE3. (B) Western blot analysis of ApoE3 in HepG2 cells incubated with added ApoE3. HepG2 cells were transfected with siRNA targeting human LRP1 (siLRP1) and non-targeting negative siRNA (siCTRL) before the rosiglitazone treatment and adding ApoE3.

Article Snippet: The lysates were subjected to SDS–PAGE using anti-human ApoE3 antibody at a 1:150 000 dilution (R&D Systems).

Techniques: Recombinant, Incubation, Western Blot, Transfection

Journal:

Article Title: Sustained Phenotypic Correction in a Mouse Model of Hypoalphalipoproteinemia with a Helper-Dependent Adenovirus Vector

doi: 10.1038/sj.gt.3302819

Figure Lengend Snippet: Structures of adenoviral vectors used for co-injection

Article Snippet: For APOE3 immunoblot, goat anti-APOE antibody (1:4000, Calbiochem) or mouse monoclonal anti-human APOE3 (1:1000, SIGNET Lab) were used.

Techniques: Control

Journal:

Article Title: Sustained Phenotypic Correction in a Mouse Model of Hypoalphalipoproteinemia with a Helper-Dependent Adenovirus Vector

doi: 10.1038/sj.gt.3302819

Figure Lengend Snippet: Co-injection of FGAd vectors suppresses hAPOA1 expression mediated by HDAd-AI vector. (a) Plasma human APOA1 levels in APOA1−/− mice treated with Ad vectors. *p<0.05 vs. PBS, **p<0.01. (b) Plasma cholesterol levels in APOA1−/− mice after intravenous injection of Ad vectors. *p<0.05 vs. PBS, **p<0.01. (c) Immunoblot analysis of transgene expression. 0.1 μl of plasma collected from mice 8 weeks after treatment with various Ad vectors was separated by SDS-PAGE, and the presence of transgene products was detected by immunoblot. Upper panel: goat anti-human APOA1 antibody which does not cross react with mouse APOA1; middle panel: goat anti-apoE antibody; lower panel: mouse monoclonal antibody against human APOE3. Lane 1: Human plasma; lane 2: APOE−/− mouse; lane 3: APOE−/− mouse treated with HDAd-E3; lane 4: APOA1−/− mouse treated with FGAd-AI + HDAd-0; lane 5: APOE−/− mouse treated with HDAd-mouse APOE; lane 6: APOA1−/− mouse treated with HDAd-AI + HDAd-0; lane 7: APOA1−/− mouse treated with HDAd-AI and HDAd-E3; lane 8: APOA1−/− mouse treated with HDAd-AI + FGAd-E3. (d) Generation of anti-hAPOA1 antibodies after intravenous co-injection of various Ad vectors. *p<0.05 (vs. day 0).

Article Snippet: For APOE3 immunoblot, goat anti-APOE antibody (1:4000, Calbiochem) or mouse monoclonal anti-human APOE3 (1:1000, SIGNET Lab) were used.

Techniques: Injection, Expressing, Plasmid Preparation, Clinical Proteomics, Western Blot, SDS Page