apoa2  (Athens Research)

Journal: Antioxidants

Article Title: Maternal Supraphysiological Hypercholesterolemia Is Accompanied by Shifts in the Composition and Anti-Atherogenic Functions of Maternal HDL along with Maternal Cardiovascular Risk Markers at Term of Pregnancy

doi: 10.3390/antiox12101804

Figure Lengend Snippet: Composition of maternal HDL from MPH and MSPH women. ( A ) Representative Western blot showing the protein abundance in HDL isolated from MPH and MSPH maternal serum. An equal quantity of protein (30 μg) was loaded for each sample. Paraoxonase 1 (PON1), apolipoprotein E (ApoE), and apolipoprotein AI (ApoAI) were determined; apolipoprotein AII (ApoAII) was used as a loading control ( n = 4 per group). Western blot quantification for specific proteins relative to ApoAII is shown in the bar graph (MPH, light blue bars, and MSPH, yellow bars). ( B ) Levels of cholesterol: total (TC), free (FC), and ester (CE) in HDL isolated from MPH (light blue bars) and MSPH (yellow bars) maternal serum. ( C ) Triglyceride levels determined in HDL isolated from MPH (light blue bar) and MSPH (yellow bar) maternal serum. n = 7 per group. Values are the mean ± S.E.M. * p < 0.05 vs. corresponding values in the MPH group.

Article Snippet: Subsequently, the proteins were transferred to polyvinylidene difluoride membranes and later probed with primary rabbit polyclonal anti-apolipoprotein B (ApoB; 1:1000 dilution), anti-apolipoprotein AI (ApoAI; 1:1000 dilution), anti-apolipoprotein AII (ApoAII; 1:1000 dilution, loading control) (Abcam, Cambridge, UK), anti-total eNOS (1:500 dilution), anti-eNOS phosphorylated at Serine 1177 (p-Ser 1177 ; 1:500 dilution), or anti-β-actin (1:2500, loading control) (Cell Signaling Technology, Danvers, MA, USA), or mouse monoclonal anti-apolipoprotein E (ApoE; 1:1000 dilution), anti-paraoxonase 1 (PON1; 1:1000 dilution) (Abcam, Cambridge, UK), or anti-ICAM-1 (1:1000 dilution) (Santa Cruz Biotechnology, Dallas, TX, USA) antibodies (18 h, 4 °C).

Techniques: Western Blot, Isolation

Journal: Antioxidants

Article Title: Maternal Supraphysiological Hypercholesterolemia Is Accompanied by Shifts in the Composition and Anti-Atherogenic Functions of Maternal HDL along with Maternal Cardiovascular Risk Markers at Term of Pregnancy

doi: 10.3390/antiox12101804

Figure Lengend Snippet: Apolipoprotein levels (g/L) in maternal serum from MPH and MSPH women. Levels of apolipoprotein AI (ApoAI), ApoB, ApoAII, ApoE, ApoCII, and ApoCIII were determined in maternal serum from MPH and MSPH pregnant women (see Methods), expressed in g/L. ApoB/ApoAI ratio was determined as a cardiovascular risk marker. * p < 0.05 vs. corresponding values in the MPH group. Values are the mean ± S.D.

Article Snippet: Subsequently, the proteins were transferred to polyvinylidene difluoride membranes and later probed with primary rabbit polyclonal anti-apolipoprotein B (ApoB; 1:1000 dilution), anti-apolipoprotein AI (ApoAI; 1:1000 dilution), anti-apolipoprotein AII (ApoAII; 1:1000 dilution, loading control) (Abcam, Cambridge, UK), anti-total eNOS (1:500 dilution), anti-eNOS phosphorylated at Serine 1177 (p-Ser 1177 ; 1:500 dilution), or anti-β-actin (1:2500, loading control) (Cell Signaling Technology, Danvers, MA, USA), or mouse monoclonal anti-apolipoprotein E (ApoE; 1:1000 dilution), anti-paraoxonase 1 (PON1; 1:1000 dilution) (Abcam, Cambridge, UK), or anti-ICAM-1 (1:1000 dilution) (Santa Cruz Biotechnology, Dallas, TX, USA) antibodies (18 h, 4 °C).

Techniques: Marker

Journal: Biological chemistry

Article Title: The redox status of cysteine thiol residues of apolipoprotein E impacts on its lipid interactions.

doi: 10.1515/hsz-2019-0414

Figure Lengend Snippet: Figure 6: Influence of apoAII on the DMPC clearance activity of apoE. Recombinant apoE2, apoE3 and apoE4 (10 mg/l) were treated with TCEP at a final concentration of 2.5 mm for 30 min at 25°C and were subsequently incubated with recombinant apoAII at final concentrations of 0 (control-2), 30 and 300 mg/l for 24 h at 37°C followed by the clearance assay. Control-1 was the apoE isoforms treated only with TCEP. Turbidity values were normalized at time zero. Time-courses for DMPC mLV clearance by apoE2 (A), apoE3 (B) and apoE4 (C) were expressed using mean values from three independent experiments. The results of the statistical analysis are summarized in Supplementary Figure 6.

Article Snippet: Recombinant apoE isoforms (apoE2, apoE3 and apoE4) were purchased from BioVision, Inc. (Milpitas, CA, USA), and recombinant apoAII was supplied by Athens Research & Technology, Inc. (Athens, GA, USA).

Techniques: Activity Assay, Recombinant, Concentration Assay, Incubation, Control

Journal: Biological chemistry

Article Title: The redox status of cysteine thiol residues of apolipoprotein E impacts on its lipid interactions.

doi: 10.1515/hsz-2019-0414

Figure Lengend Snippet: Figure 7: Influence of apoAII on the formation of apoE-DMPC liposome complexes. ApoE-DMPC liposome complexes, prepared as described in the Materials and methods section, were separated by non-denaturing PAGE using a 4–12% gradient gel, followed by detection with anti-apoE and anti-apoAII antibodies. The Stokes diameters of the complexes were determined from a calibration curve, which was constructed by plotting mobility against the corresponding Stokes diameters of a commercial size marker.

Article Snippet: Recombinant apoE isoforms (apoE2, apoE3 and apoE4) were purchased from BioVision, Inc. (Milpitas, CA, USA), and recombinant apoAII was supplied by Athens Research & Technology, Inc. (Athens, GA, USA).

Techniques: Construct, Marker