Journal: iScience

Article Title: Macrophage AMPK β1 activation by PF-06409577 reduces the inflammatory response, cholesterol synthesis, and atherosclerosis in mice.

doi: 10.1016/j.isci.2023.108269

Figure Lengend Snippet: Figure 2. PF-06409577 reduces atherosclerosis via macrophage AMPK activation (A–D) Peritoneal macrophages were isolated from AMPK b1fl/fland AMPK b1LysM mice. (A) AMPK levels and phosphorylation of ACC following treatment with PF- 06409577 (10 mM, 90 min) were detected by western blot and (B–D) quantified using ImageJ data are presented as mean G SEM, * indicates p < 0.05 and *** indicates <0.005 by unpaired t test. AMPK b1fl/fland AMPK b1 LysM mice were injected with PCSK9 AAV, fed a Western diet and treated with Vehicle or 100 mg/kg PF-06409577 by oral gavage for 6 weeks. (E and F) Representative plaque images, scale bar represents 100 mM and quantification. Data are presented as mean G SEM, # indicates p < 0.05 in overall group effect my two-way ANOVA, * indicates p < 0.05 by two-way ANOVA with Fisher’s LSD post-hoc testing.

Article Snippet: Serum PCSK9 levels were analysed using a murine PCSK9 ELISA (Cat# MPC900, R&D Systems) as per the manufacturer’s instructions.

Techniques: Activation Assay, Isolation, Phospho-proteomics, Western Blot, Injection

Journal: Frontiers in Pharmacology

Article Title: Di’ao Xinxuekang Capsule Improves the Anti-Atherosclerotic Effect of Atorvastatin by Downregulating the SREBP2/PCSK9 Signalling Pathway

doi: 10.3389/fphar.2022.857092

Figure Lengend Snippet: The primer sequences for RT-PCR assay.

Article Snippet: The mouse PCSK9 ELISA kit (batch number: E-EL-M0634c) was purchased from Elabscience Biotechnology (Wuhan, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Di’ao Xinxuekang Capsule Improves the Anti-Atherosclerotic Effect of Atorvastatin by Downregulating the SREBP2/PCSK9 Signalling Pathway

doi: 10.3389/fphar.2022.857092

Figure Lengend Snippet: Effect of DXXK, ATO, and their combination on serum lipid parameters and serum PCSK9.

Article Snippet: The mouse PCSK9 ELISA kit (batch number: E-EL-M0634c) was purchased from Elabscience Biotechnology (Wuhan, China).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: Di’ao Xinxuekang Capsule Improves the Anti-Atherosclerotic Effect of Atorvastatin by Downregulating the SREBP2/PCSK9 Signalling Pathway

doi: 10.3389/fphar.2022.857092

Figure Lengend Snippet: Effect of DXXK, ATO, and their combination on the levels of liver (A) LDLR mRNA and (B) PCSK9 mRNA, (C) LDLR and (D) PCSK9 protein expression in the liver ( n = 5); (E) the representative blot bands. Results are presented as the mean ± SD. ∗ p < 0.05 vs. ApoE −/− model group, ∗∗ p < 0.01 vs. ApoE −/− model group, ∆∆ p < 0.01 vs. ATO (1.3 mg/kg/day.) group.

Article Snippet: The mouse PCSK9 ELISA kit (batch number: E-EL-M0634c) was purchased from Elabscience Biotechnology (Wuhan, China).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Di’ao Xinxuekang Capsule Improves the Anti-Atherosclerotic Effect of Atorvastatin by Downregulating the SREBP2/PCSK9 Signalling Pathway

doi: 10.3389/fphar.2022.857092

Figure Lengend Snippet: The DXXK Improves the Anti‐atherosclerotic Effect of ATO by SREBP2/PCSK9 pathway.

Article Snippet: The mouse PCSK9 ELISA kit (batch number: E-EL-M0634c) was purchased from Elabscience Biotechnology (Wuhan, China).

Techniques:

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 increases hepatic LDLR by post-translational modification of PCSK9 in ApoE −/− mice. Eight-week-old ApoE −/− mice were fed on a high cholesterol diet (1.25% w/w) for 4 weeks and were randomized to be treated with rmSIRT1 ( n = 6) or with vehicle (PBS with 0.1% BSA) ( n = 6) for another 4 weeks. ( A ) Western blot of LDLR (120 kDa) and PCSK9 (∼62 kDa) in hepatic tissue lysates of mice ( n = 5). β-actin (45 kDa) was used as loading control. Bar graph of ELISA of ( B ) total PCSK9 and ( C ) acetylated PCSK9 in mice plasma. Statistical significance was performed using Student’s unpaired two-sample t -test. ( D , E , F ) Human hepatoma Huh7 cells [three independent triplicate experiments ( n = 3)] were treated with either 1 μmol/L of recombinant human SIRT1 (rhSIRT1) or vehicle (PBS containing 0.1% BSA) for 2 h at 37°C. The cell culture media was collected, and cells were lysed to perform ELISA to detect PCSK9. Bar graph of ELISA of ( D ) total cellular PCSK9, ( E ) total media PCSK9 and ( F ) acetylated PCSK9 in the media of cultured Huh7 cells. Grey bars represent vehicle treatment and green bars represent rmSIRT1 or rhSIRT1 treatment as indicated. Data are represented as means ± SEM. Statistical significance was performed using Student’s unpaired t -test.

Article Snippet: 20μg of PCSK9 antibody (RnD Systems) was covalently linked to beaded agarose resin (40μL) via primary amines using the Aminolink Plus immobilization kit (Pierce, catalogue number 44894).

Techniques: Modification, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Recombinant, Cell Culture

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 inhibits PCSK9-mediated LDLR degradation in hepatocytes. HuH-7 (hereafter Huh7) cells were transfected with a specific siRNA against LDLR or with non-silencing control siRNA (NS control). Assays were performed 72 h post transfection. Cells were then pre-treated with 1 μmol/L of rhSIRT1 for 2 h ( A ) Specific cellular binding of 125 I-LDL was measured at 4°C ( B ) Specific cellular association of 125 I-LDL was measured at 37°C. ( C , D , E ) Huh7 cells were co-incubated with rhPCSK9 (2 μg/mL) with rhSIRT1 (1 μmol/L) for 2 h at 37°C ( C ) Representative Western blots ( n = 3) and quantification of blot density of LDLR (130 kDa) degradation in Huh7 cells. Vinculin (140 kDa) was used as the loading control. ( D ) Specific cellular binding of 125 I-LDL was measured at 4°C ( E ) Specific cellular association of 125 I-LDL was measured at 37°C. Values are represented as means ± SEM of three independent triplicate or more experiments ( n = 3). Statistical significance was performed using one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: 20μg of PCSK9 antibody (RnD Systems) was covalently linked to beaded agarose resin (40μL) via primary amines using the Aminolink Plus immobilization kit (Pierce, catalogue number 44894).

Techniques: Transfection, Control, Binding Assay, Incubation, Western Blot, Comparison

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 inhibits PCSK9 activity through direct deacetylation in hepatocytes—specific mutation of these deacetylation sites restores PCSK9 activity. Huh7 cells were incubated with rhSIRT1 (1 μmol/L) for 2 h at 37°C. ( A ) Western blot analysis of PCSK9 immunoprecipitated Huh7 cells lysates with total PCSK9 antibody (∼62 kDa) and acetyl lysine antibody (Ac-K-PCSK9, ∼62 kDa). ( B ) Direct binding assay interaction between rhSIRT1 and rhPCSK9 using Surface Plasmon Resonance. The equilibrium dissociation constant was 34 nM. ( C ) Schematic diagram of PCSK9 domains and an overview of lysine(K) sites on PCSK9 deacetylated by SIRT1. Representative Western blot and quantification of LDLR (130 kDa) and Vinculin (140 kDa) expression in Huh7 cells expressing WT, 3KR, 3KQ mutants of PCSK9 for 72 h ( n = 3). (E )V. means empty vector ( D ) Specific cellular binding of 125 I-LDL was measured at 4°C ( E ) Specific cellular association of 125 I-LDL was measured at 37°C. ( F ) Binding of acetylated and deacetylated PCSK9 to EGF-A of LDLR using ELISA. SP—signal peptide, Pro—pro-domain, Catalytic—catalytic domain, CHRD—cysteine, histidine-rich domain. Values are represented as means ± SEM of three independent triplicate or more experiments ( n = 3). Statistical significance was performed using one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: 20μg of PCSK9 antibody (RnD Systems) was covalently linked to beaded agarose resin (40μL) via primary amines using the Aminolink Plus immobilization kit (Pierce, catalogue number 44894).

Techniques: Activity Assay, Mutagenesis, Incubation, Western Blot, Immunoprecipitation, Binding Assay, SPR Assay, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Marine drugs

Article Title: Pseurotin A Validation as a Metastatic Castration-Resistant Prostate Cancer Recurrence-Suppressing Lead via PCSK9-LDLR Axis Modulation.

doi: 10.3390/md21040215

Figure Lengend Snippet: Figure 1. Western blot analysis of PCSK9 and LDLR expression in PC cell lines and overview and validation of PCSK9 knockdown in CWR-R1ca cells. (A) PCSK9 and LDLR expression levels in diverse PC cell lines (left panel). Densitometric analysis of PCSK9 and LDLR expression levels (right panel). (B) Overview of methodology and validation of the knockdown efficacy of PCSK9 in CWR-R1ca cell transfection using shRNA-plasmid specific to PCSK9. Western blotting showed a 75% PCSK9 reduction in transfected CWR-R1ca-KD cells compared to the wild-type cells. *** p < 0.01 for statistical significance. Scanning densitometry was obtained for all blots, carried out in duplicate, and the integrated optical density of each band was normalized with the corresponding density found for β-actin in the same blot. Results are shown in the bar graphs to their respective Western blot images. Vertical bars in the graph indicate the normalized integrated optical density of bands visualized in each lane.

Article Snippet: Mice serum PCSK9 levels were measured using a Rockland ELISA kit (Rockland Immunochemicals, Inc.

Techniques: Western Blot, Expressing, Biomarker Discovery, Knockdown, Transfection, shRNA, Plasmid Preparation

Journal: Marine drugs

Article Title: Pseurotin A Validation as a Metastatic Castration-Resistant Prostate Cancer Recurrence-Suppressing Lead via PCSK9-LDLR Axis Modulation.

doi: 10.3390/md21040215

Figure Lengend Snippet: Figure 2. Effects of PS and PCSK9 knockdown on the proliferation, migration, and colony formation of CWR-R1ca cells. (A) Comparison of different PS treatments on the proliferation of wild and PCSK9- KD CWR-R1ca cells over 72 h culture period. (B) Long-term (10-day) anti-proliferative effects of PS on wild-type CWR-R1ca cells. Vertical bars indicate mean cell count ±SEM (n = 6) in MTT/proliferation assay for each treatment group over the same culture period. (C) Comparison of the migratory ability of wild and PCSK9-KD CWR-R1ca cells in wound-healing scratch assay. (D) Comparison of the antimigratory effect of PS treatments against the wild-type versus PCSK9-KD CWR-R1ca cells over 24 h. (E) Comparison of the wild-type versus PCSK9-KD CWR-R1ca cells’ colony formation ability. (F) Comparison of PS treatments’ anti-clonogenicity activity in wild-type versus PCSK9-KD CWR-R1ca cells over 12 days. Vertical bars indicate the percentage of cell migration and colony formation of wild-type and CWR-R1ca-KD cells at the experiment end calculated relative to the vehicle-treated controls (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 for statistical significance compared to VCs.

Article Snippet: Mice serum PCSK9 levels were measured using a Rockland ELISA kit (Rockland Immunochemicals, Inc.

Techniques: Knockdown, Migration, Comparison, Cell Counting, Proliferation Assay, Wound Healing Assay, Activity Assay

Journal: Marine drugs

Article Title: Pseurotin A Validation as a Metastatic Castration-Resistant Prostate Cancer Recurrence-Suppressing Lead via PCSK9-LDLR Axis Modulation.

doi: 10.3390/md21040215

Figure Lengend Snippet: Figure 3. Expression of PCSK9 in PC tissue cores. PCSK9 expression was evaluated by immuno- histochemistry in tissue microarray slide comprising 64 cases/192 PC tissue cores and 6 normal prostate cases/18 tissue cores. The protein expression was evaluated in tissues collected from PC patients at Gleason score 5 (A), GS = 7 (B), and GS = 8 (C) in addition to normal prostate tissues (D). Magnification is 400×.

Article Snippet: Mice serum PCSK9 levels were measured using a Rockland ELISA kit (Rockland Immunochemicals, Inc.

Techniques: Expressing, Immunohistochemistry, Microarray

Journal: Marine drugs

Article Title: Pseurotin A Validation as a Metastatic Castration-Resistant Prostate Cancer Recurrence-Suppressing Lead via PCSK9-LDLR Axis Modulation.

doi: 10.3390/md21040215

Figure Lengend Snippet: Figure 8. Systemic effects of HFD versus regular diet on mice serum cholesterol, LDL-C, PCSK9, and PSA levels. (A) Comparison of the HFD’s effect versus regular diet on mice mean serum total cholesterol levels. (B) Comparison of the HFD’s effect versus regular diet on mice serum LDL-C levels. (C) Comparison of the HFD’s effect versus regular diet on mice serum PCSK9 levels. (D) Comparison of the HFD’s effect versus regular diet on mice serum PSA levels. (E) Comparative of Western blotting analysis of HFD’s effect versus regular diet on the PCSK9 expression levels in CWR-R1ca cell tumors. (F) Western blot densitometric analysis of PCSK9 expression levels. Error bars represent the SEM for each experimental group. Each point is the mean of triplicate. **** p < 0.0001 for statistical significance compared to regular diet group. “ns” consider statistically not significant p > 0.5.

Article Snippet: Mice serum PCSK9 levels were measured using a Rockland ELISA kit (Rockland Immunochemicals, Inc.

Techniques: Comparison, Western Blot, Expressing

Journal: Journal of translational medicine

Article Title: Targeting PCSK9, through an innovative cVLP-based vaccine, enhanced the therapeutic activity of a cVLP-HER2 vaccine in a preclinical model of HER2-positive mammary carcinoma.

doi: 10.1186/s12967-025-06126-w

Figure Lengend Snippet: Fig. 1 Therapeutic efficacy of cVLP-PCSK9 and cVLP-HER2 vaccines as standalone or combined approach. A Experimental design; B Kaplan–Meier tumor free survival plot, n = 6; *p < 0.05; ***p < 0.01; long-rank test; C Tumor growth curves, each point represents the mean (and SEM) of all mice in each group, n = 6; D Single mice growth curves of each treated group

Article Snippet: PCSK9 protein level in mouse sera was evaluated by Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Drug discovery, Vaccines

Journal: Journal of translational medicine

Article Title: Targeting PCSK9, through an innovative cVLP-based vaccine, enhanced the therapeutic activity of a cVLP-HER2 vaccine in a preclinical model of HER2-positive mammary carcinoma.

doi: 10.1186/s12967-025-06126-w

Figure Lengend Snippet: Fig. 3 Anti-PCSK9 responses induced by cVLP-PCSK9 vaccine. A Mean (and SEM) of anti-PCSK9 antibody titers measured by ELISA represented as logarithm of Optical Density (OD) value multiplied by dilution factor of the serum sample, n = 6. Vehicle vs. cVLP-PCSK9, at least p < 0.01, from day 56; Vehicle vs. cVLP-HER2 + cVLP-PCSK9, p < 0.05, from day 98; cVLP-HER2 vs. cVLP-PCSK9, at least p < 0.05, from day 56; cVLP-PCSK9 vs. cVLP-HER2 + cVLP-PCSK9, at least p < 0.05, from day 98; cVLP-HER2 vs. cVLP-HER2 + cVLP-PCSK9, p < 0.05, day 98. All statistics were carried out by Welch t-test. B Percentage of the increase of anti-PCSK9 total antibodies after the last vaccination. Histogram reports mean (and SEM) of each group. *p < 0.05 vs Vehicle, + p < 0.05 vs cVLP-HER2, #p < 0.05 at least vs cVLP-HER2 + cVLP-PCSK9 by Welch’s t-test; C PCSK9 protein levels in mouse serum (µg/mL), each point represents the mean (and SEM) for each experimental group, n = 5–6. D Percentage of the increase of anti-PCSK9 protein inhibition after the last vaccination. Histogram reports mean (and SEM) of each group. + p < 0.05 vs cVLP-HER2, #p < 0.05 at least vs cVLP-HER2 + cVLP-PCSK9 by Welch’s t-test

Article Snippet: PCSK9 protein level in mouse sera was evaluated by Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Inhibition

Journal: Journal of translational medicine

Article Title: Targeting PCSK9, through an innovative cVLP-based vaccine, enhanced the therapeutic activity of a cVLP-HER2 vaccine in a preclinical model of HER2-positive mammary carcinoma.

doi: 10.1186/s12967-025-06126-w

Figure Lengend Snippet: Fig. 4 Inhibition of human breast cancer cell 3D agar colony growth by antibodies elicited by vaccinations. Upper panel, colony inhibition assay on BT-474 (HER2 + + + , trastuzumab sensitive) and BT-474 C5 (HER2 + + + , trastuzumab resistant), n = 2. Each bar represents the mean (and SEM) number of colonies larger than 90 µm as counted in two independent cultures with the aid of a micrometer. *p < 0.05 vs untreated, °p < 0.05 vs cVLP-PCSK9, + p < 0.05 vs cVLP-HER2, by t student’s test, unpaired. Lower panel, representative pictures of live agar colonies were acquired with an inverted microscope (dark-field, 25X)

Article Snippet: PCSK9 protein level in mouse sera was evaluated by Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Inhibition, Inverted Microscopy