Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: The total cholesterol (mg/dl) of ( A ) female ( B ) male WT and β2GPI KO mice fed with NC or HF diet, respectively. The triglyceride concentration (mg/dl) of female ( C ) and male ( D ) WT and β2GPI KO mice when fed with NC or HF diet, respectively. n = 5, * p < 0.05 by two tailed Student’s t-test. WT = wild type, mg/dl = milligrams/decilitre = WT, = β2GPI KO, NC= Normal Chow, HF= high fat, KO = Knockout

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Concentration Assay, Two Tailed Test, Knock-Out

Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: Lipoprotein separation of plasma samples from WT and β2GPI KO female and male mice fed a HF or NC diet using FPLC. ( A-D ) ( A ) Female WT and β2GPI KO mice fed a NC and ( B ) HF diet ( n = 5). ( C ) Male WT and KO mice fed NC ( n = 5) and ( D ) HF diet ( n = 4). Cholesterol levels are expressed in µg/ml. The numbers on the X axis denote FPLC fractions. VLDL eluted in fractions 13 to 21, LDL in fractions 22 to 39 and HDL in fractions 40 to 51. WT = wild type, KO = Knockout, NC = normal chow, HF= high fat = β2GPI KO = WT

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Clinical Proteomics, Knock-Out

Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: Western blot analysis of ApoE and ApoAI from individual HDL FPLC fractions comparing WT and β2GPI KO mice fed NC or HF diet. ( A ) Immunoreactivity of ApoE and ApoAI in FPLC HDL fractions from individual mice, 5 WT and 5 β2GPI KO ( A ) female mice fed NC ( B ) female mice fed a HF diet. ( C ) male mice fed NC. ( D ) male mice fed a HF diet. The ratio of Apo E to Apo AI in the HDL fractions are shown in the bar graphs below the Western blots ( A ), ( B ), ( C ), and ( D ) respectively. WT = Wild type, KO = β2GPI Knockout, NC = normal chow, HF = high fat, = WT, = β2GPI KO, ST = standard (normal plasma), Apo E = Apolipoprotein E, Apo AI = Apolipoprotein AI, MW = molecular weight, kDa = kilodaltons

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Western Blot, Knock-Out, Clinical Proteomics, Molecular Weight

Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: The plasma ApoE concentration of (A) female and (B) male WT and β2GPI KO mice when fed either a NC or HF diet. n = 5, * p < 0.05 by two-tailed Student’s t-test. NC = normal chow, HF = high fat diet, Apo E = Apolipoprotein E, ng/ml = nanograms per milliliter = WT, = β2GPI KO

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Clinical Proteomics, Concentration Assay, Two Tailed Test

Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: ( A ) β2GPI immunoreactivity in the pooled plasma lipoprotein fractions (VLDL, LDL, HDL) in female WT mice fed NC or HF diet. β2GPI reactive bands were only detected in the HDL fractions. ( B ) Quantifications of the scanned β2GPI immunoreactive bands in the HDL fractions of WT mice. Density of β2GPI in the HDL FPLC fractions from female WT mice fed a NC or HF diet. n = 5, * p < 0.05. ( C ) Immunoprecipitation of ApoE with an anti-β2GPI monoclonal antibody using plasma from female WT and β2GPI KO mice. Immunoprecipitates from WT or β2GPI KO mouse plasma was subjected to Western blot using an antibody to ApoE or β2GPI. Immunoreactive bands for β2GPI and ApoE were detected in the immunoprecipitates from WT (lane 1) but not from β2GPI KO mice (lane 2). Plasma controls consisted of WT mouse plasma demonstrating immunoreactive bands to β2GPI and ApoE (lane 3), β2GPI KO plasma did not have an immunoreactive band to β2GPI but had an immunoreactive band to ApoE (lane 4). VLDL = Very low density lipoproteins, LDL = low density lipoproteins, HDL = High density lipoproteins, KO = Knockout, HF = high fat diet, NC = normal chow, WT = Wild type

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Clinical Proteomics, Immunoprecipitation, Western Blot, Knock-Out

Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: Lipoprotein separation of human plasma samples using FPLC. ( A ) females ( B ) males. Plasma used was from 3 patients deficient in β2GPI (1 female and 2 males) and 7 age and sex matched controls (3 females and 4 males), Cholesterol levels are expressed in µg/ml. The numbers on the horizontal axis denote FPLC fractions. VLDL eluted in fractions 5 to 11, LDL in fractions 12 to 30 and HDL in fractions 31 to 46. VLDL = Very Low-density lipoprotein, LDL = low density lipoprotein, HDL = high density lipoprotein controls, β2GPI deficient patients, β2GPI = βeta-2-glycoprotein-I

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Clinical Proteomics

Journal: Biology of Sex Differences

Article Title: βeta-2 glycoprotein I is a novel regulator of Apolipoprotein E containing HDL particles in females

doi: 10.1186/s13293-025-00766-9

Figure Lengend Snippet: Effect of β2GPI deficiency on levels of ApoE in human HDL plasma samples. ( A) Western blot analysis of β2GPI, Apo E and Apo AI in pooled VLDL, LDL and HDL FPLC fractions from β2GPI deficient or control patient plasma samples. C = normal controls D = β2GPI deficient ( B ) Immunoprecipitates from one β2GPI deficient patient sample (lanes 1,3) and one normal control (lanes 2, 4) were subjected to Western blot using anti-β2GPI monoclonal antibody (lanes 1,2), or with an isotype control IgG monoclonal antibody (lanes 3, 4) and plasma from a female β2GPI deficient patient (lane 5) or normal control (lane 6). Immunoreactivity to β2GPI was not detected in any of the β2GPI deficient patient samples. In the β2GPI deficient samples ApoE immunoreactivity was only detected in the plasma sample (lane 5) but not in the immunoprecipitation samples (lane 1). Immunoreactivity for β2GPI and ApoE was detected using specific anti-human β2GPI or ApoE antibodies. ( C ) Binding of biotinylated ApoE to β2GPI and domain deletion mutants DI-IV and DII-V. % binding is expressed as a percentage of the binding of ApoE to full length β2GPI (DI-DV) which is set as 100. β2GPI = β2 glycoprotein I, ApoE = Apolipoprotein E, ApoAI = Apolipoprotein A I, MW = molecular weight, kDa = Kilodaltons

Article Snippet: Pre-cleared plasma was obtained following centrifugation for 30 s at 14,000 g. Individual plasma samples were then incubated for 14–16 h at 4°C with either sheep anti-mouse β2GPI antibody (R&D systems) (10 μg/ml) or rabbit anti-human β2GPI antibody, [ ] at 5 μg/ml final antibody concentration, [ ] followed by the addition of 20 μl of protein G-Sepharose beads.

Techniques: Clinical Proteomics, Western Blot, Control, Immunoprecipitation, Binding Assay, Molecular Weight