Journal: Acta Pharmaceutica Sinica. B

Article Title: Acteoside ameliorates hepatocyte ferroptosis and hepatic ischemia-reperfusion injury via targeting PCBP2

doi: 10.1016/j.apsb.2025.03.002

Figure Lengend Snippet: The hepatocyte specific knockout of PCBP2 interferes the therapeutic effects of ACT. (A) The particle size, (B) PDI, zeta potential and EE (%) of LNPs-si-PCBP2. (C) The stability of LNPs-si-PCBP2 in PBS and cell culture media. (D) The release behaviors of free si-RNA and LNPs-si-PCBP2. (E) Flow chart of ApoE-LNPs animal experiment. (F) The content of Fe 3+ , Fe 2+ , Cys and GSH in mouse livers. Statistical significance: ∗∗ P < 0.01, ∗∗∗ P < 0.001, compared with the sham group ( n = 6). Representative images of (G) H & E staining (scale bar = 40 μm), (H) PPB staining (scale bar = 40 μm), (I) immunofluorescence staining for PCBP2/SLC3A2/SLC7A11 (scale bar = 40 μm), (J) immunofluorescence staining for HMGB1 (scale bar = 40 μm) and (K) CD68 (scale bar = 40 μm) in mouse livers. (L) The Relative mRNA levels of slc3a2 , slc7a11 , p300 , hmgb1 and F4/80 were measured by qPCR and further normalized with hprt1 in mouse livers. Statistical significance: ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, compared with the sham group ( n = 6).

Article Snippet: The purified apolipoprotein E (ApoE) (MedChemExpress, NJ, USA) was then incubated with iminothiolane hydrochloride (2-IT) at a molar ration 1:2 and dissolved in phosphate-buffered saline at 25 °C for 90 min.

Techniques: Knock-Out, Zeta Potential Analyzer, Cell Culture, Staining, Immunofluorescence

Journal: The American journal of pathology

Article Title: The bradykinin B1 receptor regulates Aβ deposition and neuroinflammation in Tg-SwDI mice.

doi: 10.1016/j.ajpath.2013.01.021

Figure Lengend Snippet: Figure 3 B1R blockade is not associated with changes in APP processing or in levels of LRP1, APOE, and neprilysin in Tg-SwDI mice. Represen- tative blots (A) and quantitative results (B) of Western blot analysis showing that R715 admin- istration (1 mg/kg, s.c., 8 weeks) causes no changes in the levels of APP, APP C-terminal fragments C99 and C83, or APP-cleaving enzymes BACE1, ADAM10, and ADAM17, compared with vehicle-treated animals. C and D: Effect of R715 treatment on the expression of LRP1, APOE, and neprilysin. GAPDH levels were used as loading controls. The values represent means SEM (N Z 6 to 8).

Article Snippet: Western Blot Analysis Equal protein amounts were separated on a 4% to 12% SDSPAGE gradient, transferred to a nitrocellulose membrane, and incubated overnight with primary antibodies at 4 C. The following primary antibodies were used: human APP-CT20, 1741 Passos et al disintegrin and metalloproteinase domainecontaining protein (ADAM)10, ADAM17, b-site APP-cleaving enzyme 1 (BACE1; Calbiochem, San Diego, CA), glyceraldehyde-3phosphate dehydrogenase (GAPDH), B1R (Santa Cruz Biotechnology, Santa Cruz, CA), low-density lipoprotein receptorerelated protein 1 (LRP1), apolipoprotein E (APOE), neprilysin (Abcam, Cambridge, MA), phosphorylated-p65 NF-kB, liver X receptor, peroxisome proliferator-activated receptor g, and matrix metalloproteinase-9 (Cell Signaling Technology, Danvers, MA).

Techniques: Western Blot, Expressing

Journal: Journal of Lipid Research

Article Title: Amino acid residues L261, W264, F265, L268, and V269 of apolipoprotein E4 critically regulate adipose tissue metabolism

doi: 10.1016/j.jlr.2025.100958

Figure Lengend Snippet: Effects of APOE4 and APOE4mut1 on 3T3-L1 adipocyte differentiation. A: Representative Western blot analysis for APOE4, APOE4mut1 in samples of culture medium collected on the 10th day of 3T3-L1 cell differentiation as well as β-tubulin in the respective cell lysates. Samples for APOE were analysed on two separate blots. B: Semiquantitative analysis of APOE4 and APOE4mut1 levels shown in the blots of panel A, normalized for β-tubulin levels (C–E) Representative images at 10× magnification of differentiated 3T3-L1 adipocytes after Oil Red O staining. The cells were induced to differentiate after infection with: (C) AdGFP, (D) AdAPOE4, (E) AdAPOE4mut1. F: Relative absorbance of Oil Red O dye, eluted from cultures infected with recombinant adenovirus. The values were derived from the average of at least three cultures for each adenovirus used. The data were analyzed using one-way ANOVA, and the values are expressed as Mean ± SEM. G: Effect of APOE4 and APOE4mut1 on triglyceride accumulation in mature 3T3-L1 adipocytes. The values for each culture were normalized to the total protein levels of the cells present in the culture. The analysis of the results was performed using one-way ANOVA, and the values are presented as Mean ± SEM. H: mRNA expression levels of the lipogenic genes ppar γ , dgat1 , and fasn as well as ppar γ effector genes adiponectin , fabp4 , fabp5 , and lpl in differentiated 3T3-L1 adipocytes expressing APOE4 and APOE4mut1. The results were normalized to the expression levels of the housekeeping rps18 gene. The analysis was performed using two-way ANOVA, and the values are presented as Mean ± SEM. ∗∗ = P < 0.005 and ∗∗∗ = P < 0.0005, n = 3 per group. Unless indicated by a horizontal line between two groups, statistical significance refers to differences among all test groups.

Article Snippet: For the semiquantitative measurement of murine and human protein levels in the plasma and the lipoprotein fractions of the mice, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was performed, followed by Western blot, using the following antibodies: Rabbit anti-Human and anti-Mouse APOE (D7I9N, cat# 10197, Cell Signaling), Goat anti APOB-48/100 HRP (cat# K34005G, Meridian Life Science), Rabbit anti-Mouse β-tubulin(cat# # 2146, Cell Signaling), Rabbit anti-Mouse Cytc (cat# 4272, Cell Signaling), Rabbit anti-Mouse Ucp1 (E9Z2V, cat#72298, Cell Signaling), Rabbit anti-Mouse Cox4 (cat# 4844, Cell Signaling), and Goat anti-rabbit IgG-HRP (cat#7074, Cell Signaling).

Techniques: Western Blot, Cell Differentiation, Staining, Infection, Recombinant, Derivative Assay, Expressing

Journal: Journal of Lipid Research

Article Title: Amino acid residues L261, W264, F265, L268, and V269 of apolipoprotein E4 critically regulate adipose tissue metabolism

doi: 10.1016/j.jlr.2025.100958

Figure Lengend Snippet: Plasma lipid and glucose levels of C57BL/6 mice fed high-fat diet for 8 weeks and then infected with the adenoviruses AdGFP, AdAPOE4, and AdAPOE4mut1. A: Representative Western blot of human and murine APOE five days after infection of mice with the adenoviruses, (B) plasma total cholesterol, (C) plasma total triglyceride, and (D) plasma glucose levels five days after infection of mice with the adenoviruses. The results were analyzed using one-way ANOVA. Values are expressed as Mean ± SEM. ∗ = P < 0.05, ∗∗ = P < 0.005 and ∗∗∗ = P < 0.0005, n = 3 per group.

Article Snippet: For the semiquantitative measurement of murine and human protein levels in the plasma and the lipoprotein fractions of the mice, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was performed, followed by Western blot, using the following antibodies: Rabbit anti-Human and anti-Mouse APOE (D7I9N, cat# 10197, Cell Signaling), Goat anti APOB-48/100 HRP (cat# K34005G, Meridian Life Science), Rabbit anti-Mouse β-tubulin(cat# # 2146, Cell Signaling), Rabbit anti-Mouse Cytc (cat# 4272, Cell Signaling), Rabbit anti-Mouse Ucp1 (E9Z2V, cat#72298, Cell Signaling), Rabbit anti-Mouse Cox4 (cat# 4844, Cell Signaling), and Goat anti-rabbit IgG-HRP (cat#7074, Cell Signaling).

Techniques: Clinical Proteomics, Infection, Western Blot

Journal: Journal of Lipid Research

Article Title: Amino acid residues L261, W264, F265, L268, and V269 of apolipoprotein E4 critically regulate adipose tissue metabolism

doi: 10.1016/j.jlr.2025.100958

Figure Lengend Snippet: Plasma lipoprotein lipid levels of C57BL/6 mice fed high-fat diet for 8 weeks and then infected with the adenoviruses AdGFP, AdAPOE4, and AdAPOE4mut1. Lipoproteins were fractionated by KBr density gradient ultracentrifugation. A, C–E: Total cholesterol and (F–I) triglyceride levels of lipoprotein fractions 5 days after infection of mice with the adenoviruses. Panel B shows the murine APOE, human APOE4 and APOE4mut1 distribution among lipoprotein fractions. The results were analyzed using one-way ANOVA, and values are expressed as Mean ± SEM. ∗∗∗ = P < 0.0005, n = 3 per group. Statistical significance refers to differences among all test groups.

Article Snippet: For the semiquantitative measurement of murine and human protein levels in the plasma and the lipoprotein fractions of the mice, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was performed, followed by Western blot, using the following antibodies: Rabbit anti-Human and anti-Mouse APOE (D7I9N, cat# 10197, Cell Signaling), Goat anti APOB-48/100 HRP (cat# K34005G, Meridian Life Science), Rabbit anti-Mouse β-tubulin(cat# # 2146, Cell Signaling), Rabbit anti-Mouse Cytc (cat# 4272, Cell Signaling), Rabbit anti-Mouse Ucp1 (E9Z2V, cat#72298, Cell Signaling), Rabbit anti-Mouse Cox4 (cat# 4844, Cell Signaling), and Goat anti-rabbit IgG-HRP (cat#7074, Cell Signaling).

Techniques: Clinical Proteomics, Infection

Journal: Journal of Biological Chemistry

Article Title: ATP-binding Cassette Transporter A1 Mediates the Beneficial Effects of the Liver X Receptor Agonist GW3965 on Object Recognition Memory and Amyloid Burden in Amyloid Precursor Protein/Presenilin 1 Mice*

doi: 10.1074/jbc.m110.108100

Figure Lengend Snippet: FIGURE 2. High dose GW3965 is required for elevated apoE levels in tissue. Cortical and hippocampal extracts from APP/PS1 untreated (: white), APP/PS1 treated (: dark gray), APP/PS1 ABCA1/ untreated (: light gray), and APP/PS1 ABCA1/ treated (: black) mice were immunoblotted for apoE. Left panels are representative blots using undiluted samples from individual mice from the following cohorts: APP/PS1 untreated control: n 7 (cortex), n 8 (hippocampus); APP/PS1 prophylactic: n 8 (cortex) n 8 (hippocam- pus); APP/PS1 low dose therapeutic: n 8 (cortex), n 8 (hippocampus); APP/PS1 high dose therapeutic: n 10 (cortex), n 9 (hippocampus); ABCA1/ APP/PS1 untreated control: n 4 (cortex), n 4 (hippocampus); ABCA1/ APP/PS1 prophylactic: n 8 (cortex) n 8 (hippocampus): ABCA1/ APP/PS1 low dose therapeu- tic: n 7 (cortex), n 6 (hippocampus); ABCA1/ APP/PS1 high dose therapeutic: n 4 (cortex), n 4 (hippocampus). Samples from each mouse were blotted at least twice, normalized to actin, and expressed as fold-difference compared with apoE levels in untreated APP/PS1 controls. Data represent mean S.E., where * represents p 0.05 and ** represents p 0.01 by one-way analysis of variance followed by Tukey post test.

Article Snippet: Western Blot Analysis of ABCA1, ApoE, APP, and APP-CTF— For analysis ofABCA1, apoE, andAPP, 50–100 g of carbonate lysate was electrophoresed through 10% SDS-polyacrylamide gels, transferred to PVDFmembrane (Millipore), and immunodetected using a monoclonal anti-ABCA1 antibody, AC10 (1:1000, a kind gift from Dr. M. R. Hayden), a murine-specific apoE antibody (1:1000, Santa Cruz Biotechnology), an antiAPP antibody that recognizes both murine and human APP (clone 22C11, 1:2000, Chemicon), or an anti-actin antibody (1:5000, Chemicon) as a loading control.

Techniques: Control

Journal: Journal of Biological Chemistry

Article Title: ATP-binding Cassette Transporter A1 Mediates the Beneficial Effects of the Liver X Receptor Agonist GW3965 on Object Recognition Memory and Amyloid Burden in Amyloid Precursor Protein/Presenilin 1 Mice*

doi: 10.1074/jbc.m110.108100

Figure Lengend Snippet: FIGURE 3. ABCA1 is required for increased CSF apoE levels in response to GW3965. Equal volumes of CSF from individual mice from untreated control (C), prophylactic (P), low dose therapeutic (T), and high dose therapeutic (TH) groups of APP/PS1 and APP/PS1 ABCA1/ mice were pooled, separated by 6% native PAGE, and immunoblotted for apoE and albumin. Stokes diameter markers are shown on the right.

Article Snippet: Western Blot Analysis of ABCA1, ApoE, APP, and APP-CTF— For analysis ofABCA1, apoE, andAPP, 50–100 g of carbonate lysate was electrophoresed through 10% SDS-polyacrylamide gels, transferred to PVDFmembrane (Millipore), and immunodetected using a monoclonal anti-ABCA1 antibody, AC10 (1:1000, a kind gift from Dr. M. R. Hayden), a murine-specific apoE antibody (1:1000, Santa Cruz Biotechnology), an antiAPP antibody that recognizes both murine and human APP (clone 22C11, 1:2000, Chemicon), or an anti-actin antibody (1:5000, Chemicon) as a loading control.

Techniques: Control, Clear Native PAGE