Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Identification of a peptide inhibitor disrupting the PCSK9-LDLR interaction via pharmacophore-based virtual screening, molecular dynamics simulations and in vitro/vivo evaluation

doi: 10.1080/14756366.2025.2610849

Figure Lengend Snippet: The pharmacophore model of the LDLR-binding domain in PCSK9 was established using MOE. Pharmacophore features include: F1, an aromatic feature; F2 and F4, hydrogen-bond acceptor features; F3, a hydrogen-bond donor feature.

Article Snippet: The supernatants were used to determine hepatic LDLR protein levels using a mouse LDLR ELISA kit (R&D Systems).

Techniques: Binding Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Identification of a peptide inhibitor disrupting the PCSK9-LDLR interaction via pharmacophore-based virtual screening, molecular dynamics simulations and in vitro/vivo evaluation

doi: 10.1080/14756366.2025.2610849

Figure Lengend Snippet: The 2D interaction plots for TPPs 1–5 and the LDLR-binding domain in PCSK9 using the LigPlot. The hydrogen bonds are shown in green.

Article Snippet: The supernatants were used to determine hepatic LDLR protein levels using a mouse LDLR ELISA kit (R&D Systems).

Techniques: Binding Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Identification of a peptide inhibitor disrupting the PCSK9-LDLR interaction via pharmacophore-based virtual screening, molecular dynamics simulations and in vitro/vivo evaluation

doi: 10.1080/14756366.2025.2610849

Figure Lengend Snippet: The 3D interaction plots for TPPs 1–5 and the LDLR-binding domain in PCSK9. (A, B) TPP-1 are colour-coded by yellow. (C, D) TPP-2, purple; (E, F) TPP-3, orange; (G, H) TPP-4, cyan; (I, J) TPP-5, green. The LDLR-binding domain residues of PCSK9 are displayed as sticks: nitrogen atoms are blue, oxygen atoms are red, and other residues are silver.

Article Snippet: The supernatants were used to determine hepatic LDLR protein levels using a mouse LDLR ELISA kit (R&D Systems).

Techniques: Binding Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Identification of a peptide inhibitor disrupting the PCSK9-LDLR interaction via pharmacophore-based virtual screening, molecular dynamics simulations and in vitro/vivo evaluation

doi: 10.1080/14756366.2025.2610849

Figure Lengend Snippet: The surface LDLR expression in HepG2 cells. (A) HepG2 cells treated with TPPs 1–5, Pep2-8, and Evolocumab. (B) HepG2 cells treated with TPP-4 at different concentrations. *** P < 0.001 vs Pep2-8.

Article Snippet: The supernatants were used to determine hepatic LDLR protein levels using a mouse LDLR ELISA kit (R&D Systems).

Techniques: Expressing

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Identification of a peptide inhibitor disrupting the PCSK9-LDLR interaction via pharmacophore-based virtual screening, molecular dynamics simulations and in vitro/vivo evaluation

doi: 10.1080/14756366.2025.2610849

Figure Lengend Snippet: (A) Liver LDLR levels in mice treated with TPP-4 and Evolocumab. (B) Plasma total cholesterol levels in mice treated with TPP-4 and Evolocumab.

Article Snippet: The supernatants were used to determine hepatic LDLR protein levels using a mouse LDLR ELISA kit (R&D Systems).

Techniques: Clinical Proteomics

Journal: Nucleic Acids Research

Article Title: Antisense oligonucleotides targeting translation inhibitory elements in 5′ UTRs can selectively increase protein levels

doi: 10.1093/nar/gkx632

Figure Lengend Snippet: Translation of LDLR protein is enhanced by targeting the predicted structure using an ASO. ( A ) The potential effect of an ASO on the predicted structure is depicted. ( B ) ELISA analyses for LDLR protein levels in HEK293 cells transfected for 15 h with XL824, which targets the 5′ side of the predicted stem. ( C ) qRT-PCR quantification of LDLR mRNA in cells treated with XL824. ( D ) ELISA analyses for LDLR protein in HEK293 cells transfected for 15 h with ASO814923, targeting the 3′ side of the potential stem or an ASO XL506 targeting a potential uORF. ASO concentration 0 indicates mock transfection. ( E ) Western analyses for LDLR protein in HEK293 cells treated with 30 nM ASO814923 for 10 h. Levels were normalized to those of GAPDH. ( F ) qRT-PCR quantification of LDLR mRNA levels in HEK293 cells treated with the ASO. ( G ) Autoradiography of 35 S-methionine-labeled nascent LDLR protein isolated by immunoprecipitation from cells treated with ASO814923 (+) or mock-treated (–) for 7 h. An aliquot of cell lysate used for immunoprecipitation was analyzed by SDS-PAGE, and labeled nascent proteins were visualized by autoradiography (right panel). The level of nascent LDLR protein was normalized to signal from band marked by an arrow, and relative levels are given below the lanes. ( H ) qRT-PCR quantification of LDLR mRNA co-immunoprecipitated using an anti-eIF4A antibody or control IgG from mock transfected HEK293 cells (UTC) or cells treated with the ASO814923 for 10 h. 7SL RNA and RNASEH1 mRNA were quantified as controls. ( I ) LDL uptake in HEK293 cells treated with 30 nM ASO814923 or 30 nM control ASO812662 for 15 h. ( J ) qRT-PCR for LDLR mRNA levels in HEK293 cells treated with siRNA targeting luc or LDLR for 4 h, followed by treatment with or without ASO814923 (40 nM) for an additional 10 h. ( K ) ELISA analyses for LDLR protein levels in HEK293 cells treated with siRNAs, followed by treatment with ASO814923. ( L ) LDL uptake in HEK293 cells treated with siRNAs for 4 h, followed by treatment of the ASO for 10 h, and LDL uptake analysis. The error bars are standard deviations from three experiments. P -values were calculated based on unpaired t-test. NS, not significant. ** P < 0.01; *** P < 0.001 and **** P < 0.0001.

Article Snippet: Around 10 μg total protein was analyzed using an ELISA kit (DLDLR0, R&D Systems) for human LDLR based on the manufacturer's protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Transfection, Quantitative RT-PCR, Concentration Assay, Western Blot, Autoradiography, Labeling, Isolation, Immunoprecipitation, SDS Page, Control

Journal: Nucleic Acids Research

Article Title: Antisense oligonucleotides targeting translation inhibitory elements in 5′ UTRs can selectively increase protein levels

doi: 10.1093/nar/gkx632

Figure Lengend Snippet: ASOs with different modifications can increase LDLR protein levels. ( A ) ELISA analysis for LDLR protein levels in HeLa cells treated with ASO842196 for 15 h at different concentrations. ( B ) ELISA analyses for LDLR protein levels in HEK293 cells transfected with ASO842196, ASO842197, or ASO842206 at indicated concentrations for 12 h. Means ± standard deviations of three independent experiments are plotted. P -values were calculated based on unpaired t -test. *** P < 0.001; **** P < 0.0001.

Article Snippet: Around 10 μg total protein was analyzed using an ELISA kit (DLDLR0, R&D Systems) for human LDLR based on the manufacturer's protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Transfection