Journal: Diabetes & metabolism

Article Title: Large triglyceride-rich lipoproteins from fasting patients with type 2 diabetes activate platelets.

doi: 10.1016/j.diabet.2019.03.002

Figure Lengend Snippet: Fig. 2. Large triglyceride-rich lipoproteins (TGRL) from type 2 diabetes (T2D) patients increase thrombin-stimulated platelet aggregation and thromboxane B2 (TxB2) concentrations via activation of the arachidonic acid signalling pathway: (A) platelets from healthy blood donors (HBD) were incubated with/without T2D large TGRL (25 mg proteins/mL), followed by stimulation with thrombin (0.013 U/mL 0.001) and agitated at 1000 rpm for 4 min. Results are the mean SEM percentage of platelets stimulated by thrombin (n = 6; * P < 0.05); (B) platelets from HBD were incubated at 37 8C for 30 min with/without T2D large TGRL (25 mg proteins/mL) and stimulated with thrombin (0.1 U/ mL) for 1 h at 37 8C while undergoing slow agitation (n = 13). Results are expressed as mean SEM TxB2 concentrations (pmol/109 platelets). Platelets + thrombin vs. platelets alone: + 1453%, * P < 0.05; platelets + T2D large TGRL + thrombin vs. platelets + thrombin: + 37%, * P < 0.05; (C, D) platelets from healthy donors were incubated at 37 8C with/ without thrombin (0.09 0.01 U/mL) while undergoing slow agitation. Platelets were incubated with 25 mg of proteins/mL of T2D large TGRL and thrombin under the same test conditions. Phosphorylation of p38 MAPK and cPLA2 was measured after sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) separation and immunoblotting with anti-phospho-p38 MAPK and anti-phospho-cPLA2 antibodies (n = 6 and n = 3, respectively). Results are the mean percentage of platelets stimulated by thrombin. Platelets + thrombin vs. platelets alone: * P < 0.05; platelets + T2D large TGRL + thrombin vs. platelets + thrombin: * P < 0.05 and representative phospho-p38 MAPK immunoblot; platelets + thrombin vs. platelets alone: * P < 0.05; platelets + T2D large TGRL + thrombin vs. platelets + thrombin: * P < 0.05 and representative phospho-cPLA2 immunoblot.

Article Snippet: These membranes were blocked with 5% skimmed milk and incubated with either 1:2500 anti-phosphop38 MAPK or 1:1000 anti-phospho-cPLA2 polyclonal antibodies (Cell Signaling Technology, Danvers, MA, USA), then washed and incubated with 1:5000 or 1:2000 goat anti-rabbit horseradish peroxidase conjugate.

Techniques: Activation Assay, Incubation, Phospho-proteomics, Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot

Journal: Reproductive Sciences

Article Title: The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

doi: 10.1007/s43032-021-00691-3

Figure Lengend Snippet: Gene-related information

Article Snippet: Primary antibodies: mouse anti–human ABCA1 polyclonal antibody (Abcam; diluted 1:3000); rabbit anti–human LDLR polyclonal antibody (PTG; diluted 1:1000); rabbit anti–human SCARB1 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human SULT2B1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CYP19A1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CRLS1 polyclonal antibody (PTG; diluted 1:1000); mouse anti–human LPCAT3 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human PLA2G4A polyclonal antibody (Abclonal; diluted 1:1000).

Techniques:

Journal: Reproductive Sciences

Article Title: The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

doi: 10.1007/s43032-021-00691-3

Figure Lengend Snippet: Protein expression in CCs and MGCs. Primary antibodies: mouse anti–human ABCA1 polyclonal antibody (Abcam; diluted 1:3000); rabbit anti–human LDLR polyclonal antibody (PTG; diluted 1:1000); rabbit anti–human SCARB1 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human SULT2B1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CYP19A1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CRLS1 polyclonal antibody (PTG; diluted 1:1000); mouse anti–human LPCAT3 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human PLA2G4A polyclonal antibody (Abclonal; diluted 1:1000). Secondary antibodies: goat anti-mouse IgG conjugated with horseradish peroxidase (HRP) diluted 1:3000 in TTBS for ABCA1 and LPCAT3 or goat anti–rabbit IgG conjugated with HRP diluted 1:3000 in TTBS for LDLR, SCARB1, SULT2B1, CYP19A1, CRLS1, and PLA2G4A. The expression of mouse anti–human ACTIN polyclonal antibody (Servicebio; diluted 1:3000) was used as an internal reference. A shows the protein expression of ABCA1, LDLR, SCARB1, SULT2B1, and CYP19A1; B shows the protein expression of CRLS1, LPCAT3, and PLA2G4A

Article Snippet: Primary antibodies: mouse anti–human ABCA1 polyclonal antibody (Abcam; diluted 1:3000); rabbit anti–human LDLR polyclonal antibody (PTG; diluted 1:1000); rabbit anti–human SCARB1 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human SULT2B1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CYP19A1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CRLS1 polyclonal antibody (PTG; diluted 1:1000); mouse anti–human LPCAT3 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human PLA2G4A polyclonal antibody (Abclonal; diluted 1:1000).

Techniques: Expressing

Journal: Reproductive Sciences

Article Title: The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

doi: 10.1007/s43032-021-00691-3

Figure Lengend Snippet: Gene expression of MGCs and CCs was evaluated by qPCR. A shows the gene expression of ABCA1, ABCG1, LDLR, and SCARB1; B shows the gene expression of SULT2B1, STS, and CYP19A1. In A and B , the gene expression of MGCs is considered to be 1; C shows the gene expression of LPCAT3 and PLA2G4A; D shows the gene expression of PTPMT1 and CRLS1. In C and D , the gene expression of MGCs is considered to be 1. * P < 0.05, ** P < 0.01, Student’s t -test

Article Snippet: Primary antibodies: mouse anti–human ABCA1 polyclonal antibody (Abcam; diluted 1:3000); rabbit anti–human LDLR polyclonal antibody (PTG; diluted 1:1000); rabbit anti–human SCARB1 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human SULT2B1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CYP19A1 polyclonal antibody (Abclonal; diluted 1:1000); rabbit anti–human CRLS1 polyclonal antibody (PTG; diluted 1:1000); mouse anti–human LPCAT3 polyclonal antibody (Abcam; diluted 1:1000); rabbit anti–human PLA2G4A polyclonal antibody (Abclonal; diluted 1:1000).

Techniques: Expressing