Journal: JCI Insight

Article Title: Platelets impair the resolution of inflammation in atherosclerotic plaques in insulin-resistant mice after lipid lowering

doi: 10.1172/jci.insight.193593

Figure Lengend Snippet: ( A ) Study design. Eight-week-old Ldlr –/– male mice fed a high-fat, high-cholesterol (HFHC) diet for the whole duration of the study (23 weeks). After the atherosclerosis progression period (obese baseline, Ob), mice were injected with ApoB ASO to lower lipids (lipid-lowering period). All lipid-lowered mice were split into 2 groups, injected either with isotype control antibody (IgG) or platelet depletion antibody (αCD42b) every 3 days for a total of 3 weeks. ( B ) Platelet counts in circulating blood of Ob, IgG, and αCD42b 3 days before harvest. ( C ) Blood glucose measurements (lean n = 16, Ob n = 47) and ( D ) area of curve (AOC) quantification of glucose tolerance tests (GTTs) from lean, Ob, IgG, and αCD42b mice after 16 weeks of HFHC diet during atherosclerosis progression. ( E ) Blood glucose measurements (Ob n = 3, IgG n = 4, αCD42b n = 4) and ( F ) AOC quantification of GTTs from Ob, IgG, and αCD42b mice after 22 weeks of HFHC diet during atherosclerosis resolution. Mean fluorescent intensity (MFI) of platelet ( G ) JonA and ( H ) P-selectin in nonstimulated (NS) samples and upon 100 μM PAR4-AP agonist stimulation of lean and Ob circulating blood samples. ( I ) Mean platelet volume (MPV) from circulating blood of lean, Ob, and IgG-treated mice, 3 days before harvest. In C and E , error bars represent SD. Data were analyzed by Kruskal-Wallis with Dunn’s post hoc test ( B and F ), unpaired Mann-Whitney test ( C , D , and H ), unpaired Student’s t test ( G ), or ordinary 1-way ANOVA with Tukey’s multiple-comparison test ( I ). P values are shown in graphs.

Article Snippet: The whole blood was stained with anti–mouse JonA-PE (M023-2, Emfret Analytics) and anti–mouse P-selectin Alexa Fluor 647 (563674, BD Biosciences) and then treated with PAR4-AP platelet agonist (100 μM; MedChemExpress) for 15 minutes at room temperature.

Techniques: Injection, Control, MANN-WHITNEY, Comparison

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 4. Screening of key prognostic genes. (A) Protein–protein interaction (PPI) analysis. (B) KEGG analysis. (C) Expression of F2RL3 between tumor and normal tissues. (D) Expression of F2RL3 in GC stages. (E) Kaplan-Meier (K-M) analysis of F2RL3.

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Expressing

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 5. F2RL3 expression level in human GC tissue. (A) F2RL3 expression detected by Western blotting (n = 6). (B) Bar chart

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Expressing, Western Blot

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 6. F2RL3 expression level in human GC cells. (A) F2RL3 expression detected by Western blotting. (B) Bar chart showing F2RL3

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Expressing, Western Blot

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 7. Effects of F2RL3 knockdown on the EMT of GC cells. (A) F2RL3 expression detected by Western blotting. (B) Bar chart

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Knockdown, Expressing, Western Blot

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 8. Effects of F2RL3 expression knockdown on angiogenesis. (A) The activity of Human Umbilical Vein Endothelial Cells

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Expressing, Knockdown, Activity Assay

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 9. F2RL3 regulates the expression of Rap1/MAPK signaling pathway in GC cells. (A) Expression of proteins related to F2RL3

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Expressing

Journal: Frontiers in bioscience (Landmark edition)

Article Title: F2RL3 Regulates Epithelial-Mesenchymal Transition and Angiogenesis in Gastric Cancer through the Rap1/MAPK Signaling Pathway.

doi: 10.31083/j.fbl2905177

Figure Lengend Snippet: Fig. 10. Effects of the Rap1/MAPK signaling pathway on the EMT of GC cells. (A) Expression of F2RL3, E-cadherin, vimentin, and Snail detected by Western blotting. (B) Bar chart showing the expression of F2RL3, vimentin, E-cadherin, and Snail. (C) Activity

Article Snippet: Then the sections were incubated with F2RL3 primary antibody (#25306-1-AP; Proteintech, Rosemont, IL, USA), followed by incubation with secondary antibody.

Techniques: Expressing, Western Blot, Activity Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: PAR4 Inhibition Reduces Coronary Artery Atherosclerosis and Myocardial Fibrosis in SR-B1/LDLR Double Knockout Mice

doi: 10.1161/ATVBAHA.123.319767

Figure Lengend Snippet: RAG8 treatment impairs PAR (protease-activated receptor) 4–mediated platelet activation. A , SR-B1 (scavenger receptor class B type I)/LDLR (low-density lipoprotein receptor) double knockout (DKO) mice fed the high-fat, high-cholesterol diet containing cholate (HFCC) for 20 days and treated with the RAG8 or the inactive control SRQ8 pepducins were anesthetized, and their tails were transected across the vein ≈0.5 cm from the tip and placed in a cuvette with warm saline. The time to cessation of bleeding was monitored. Each data point represents an individual mouse (n=12, 14). B and C , SR-B1/LDLR DKO mice were fed the HFCC for 14 days and treated with the RAG8 or the inactive control SRQ8 pepducins (n=4 each). Platelets were prepared from blood collected retro-orbitally from anesthetized mice. Platelet activation was measured by treatment of platelets with either thrombin or a synthetic PAR4-AP (PAR4 agonist peptide) by flow cytometry after immunostaining with either ( B ) a FITC (fluorescein isothiocyanate)-labeled Wug-E9 antibody (for P-selectin) or ( C ) a PE (phycoerythrin)-labeled JON/A antibody for active GP IIb/IIIa (glycoprotein). Resting platelet activation levels do not differ between control SRQ8- and RAG8-treated SR-B1/LDLR DKO mice. Units are expressed as median fluorescence intensity (MFI). Bars represent means, and error bars represent SEMs. Data in A passed the normality test and were analyzed by Student t test. Data in B and C were analyzed by 2-way ANOVA with Sidak multiple comparisons post hoc test. **** P <0.0001.

Article Snippet: For pepducin-treated mouse groups, platelets (1×10 6 washed mouse platelets per reaction) were resting or activated with 1 U/mL α-thrombin (HT 1002A; Enzyme Research Laboratories, South Bend, IN) or 300 μM PAR4 agonist peptide (AYPGKF-NH 2 ; custom synthesized by Genscript, Piscataway, NJ) in the presence of 2 mM CaCl 2 .

Techniques: Activation Assay, Double Knockout, Control, Saline, Flow Cytometry, Immunostaining, Labeling, Fluorescence