gpc2 his  (R&D Systems)

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: A: Brachial artery flow-mediated dilation (FMD, expressed as percent change in arterial diameter), plasma neuraminidase and ADAM17 activities (expressed as fold difference from healthy participants (males, n=12; females, n=18; open squares and triangles, respectively) and participants with T2D (males, n=26; females, n= 21; solid squares and triangles, respectively). B: FMD and sodium nitroprusside (SNP)-induced vasodilation in mouse-isolated femoral arteries (n=5-6/group). Arteries were preconstricted with phenylephrine. C: Representative images (left panel) and quantification (right panel) of endothelial glypican-1 content on the luminal surface of the endothelium in isolated femoral arteries from wild-type (WT) and db/db mice (n=8/group). Data are presented as means ± SE. Mann-Whitney test (A). Two-way repeated-measures ANOVA (B). Two-tailed unpaired t test (C). * P <0.05 vs. healthy participants (A) or WT mice (B-C).

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Shear, Clinical Proteomics, Isolation, MANN-WHITNEY, Two Tailed Test

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: Endothelial cells were exposed to neuraminidase (Neu, 100 mU/mL) vs. vehicle for 1 hr. A: PS externalization (n=20-24/condition); B: ADAM17 activity (n=12/condition); C: shedding of glypican-1 (cell culture supernatant relative to lysate, n=12/condition, including representative blots. Data are presented as means ± SE. Two-tailed unpaired t test (A-C). *P <0.05 vs. respective controls.

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Activity Assay, Cell Culture, Two Tailed Test

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: Endothelial cells were exposed to Neu (100 mU/mL) for 1 hr with or without an ANO6-i (A6-001,10 μM). ANO6-i was added 1 hr before Neu. A: PS externalization (n=9/condition); B: ADAM17 activity (n=12/condition); C: shedding of glypican-1 (cell culture supernatant relative to lysate, n=7-8/condition, including representative blots) in endothelial cells. Data are presented as means ± SE. Two-tailed unpaired t test (A-C). *P <0.05 vs. respective controls.

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Inhibition, Activity Assay, Cell Culture, Two Tailed Test

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: Endothelial cells were exposed to Neu (100 mU/mL) for 1 hr with or without an ADAM17 inhibitor (TAPI-0, 50 μM). TAPI-0 was added 1 hr before Neu. A: PS externalization (n=11-12/condition); B: ADAM17 activity (n=11-23/condition); C: shedding of glypican-1 (cell culture supernatant relative to lysate, n=11-12/condition, including representative blots) in endothelial cells. Data are presented as means ± SE. Two-tailed unpaired t test (A-C). *P <0.05 vs. respective controls.

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Inhibition, Activity Assay, Cell Culture, Two Tailed Test

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: A-B: ADAM17 expression (n=12/condition, including representative blots), ADAM17 activity (n=7-24/condition), and shedding of glypican-1 (n=27-32/condition, including representative blots). C-D: p-eNOS Ser1177 relative to total protein (n=15/condition, including representative blots) and total eNOS (n=4/condition, including representative images, scale bar=40 μm) in response to shear stress (15 dynes/cm 2 , 2 hr and 24 hr, respectively) in cells infected with control or ADAM17-overexpressing adenovirus (ADAM17-OE) for 24 hr. E: siRNA-mediated silencing of glypican-1 impairs the upregulation of eNOS in response to 24 hr shear stress (n=4/condition, including representative images, scale bar=40 μm). The inset depicts the downregulation of surface glypican-1 via siRNA. F: Intraluminal exposure of human omental arteries to active ADAM17-r protein (1 ug/mL) impairs FMD (n=9/condition). The response to SNP remained unchanged (data published in ( 13 )). G: Cleavage of recombinant glypican-1 protein (glypican-1-r, 33.3 μg/mL) by recombinant ADAM17 protein (ADAM17-r, 166.7 μg/mL) in the presence or absence of TAPI-0 in a cell-free based assay following an 8-hr incubation at 37°C (n=5-6/condition, including representative total protein gel). H: Identified ADAM17-dependent cleavage site of glypican-1 based on proteomic analysis performed at the University of Missouri Charles W. Gehrke Proteomics Center. Illustration of the iceLogo cleavage site was adapted with permission from work by Tucher et al. ( 32 ). Data are presented as means ± SE. Two-tailed unpaired t test (A and B). One-way ANOVA (A, G). Two-way ANOVA without (C-E) and with repeated measurements (F). *, # P <0.05 vs. respective controls. § P <0.05 vs. control with shear. $ P <0.05 Main effect of glypican-1 knockdown (E).

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Shear, Isolation, Expressing, Activity Assay, Infection, Control, Recombinant, Cell-Free Assay, Incubation, Two Tailed Test, Knockdown

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: A: Pulmonary endothelial cells (control, n=12; ADAM17-KO, n=14) were assessed for ADAM17 content, which was reduced in the knockout mice. No differences were observed between groups for B : body weight (control, n=28; ADAM17-KO, n=39), C : blood glucose during a glucose tolerance test (control, n=29; ADAM17-KO, n=39), or D : systolic blood pressure (BP) (control, n=28; ADAM17-KO, n=38). ADAM17-KO mice had greater FMD in isolated mesenteric arteries compared to genetic controls. No differences in SNP-induced vasodilation were noted. Vessels were preconstricted with phenylephrine (males, n=31; females, n=20). C. Mice that were ADAM17-KO also had greater glypican-1 content compared to genetic controls (control, n=8; ADAM17-KO, n=13). Data are presented as means ± SE. Two-tailed unpaired t test (A and F). Two-way repeated-measures ANOVA (B). *P <0.05 vs. respective controls.

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Isolation, Western Blot, Control, Knock-Out, Two Tailed Test

Journal: American journal of physiology. Cell physiology

Article Title: ADAM17-induced shedding of glypican-1 as a mechanism of impaired endothelial shear stress mechanotransduction

doi: 10.1152/ajpcell.00794.2025

Figure Lengend Snippet: Neuraminidase, which is elevated in the circulation of individuals with type 2 diabetes, induces an increase in endothelial intracellular Ca 2+ (1). This Ca 2+ increase is likely the stimulus for activation of Anoctamin-6 (ANO6), which flips phosphatidylserine (PS) to the outer leaflet of the plasma membrane (2). The PS head group is negatively charged. Loss of plasma membrane asymmetry by flipped PS causes its interaction with the membrane proximal domain of ADAM17, which presents a cationic PS-binding motif. This interaction causes ADAM17 to bend to the appropriate position for substrate shedding, including mechanosensitive structures such as glypican-1 (3). Loss of glypican-1 likely contributes to impaired shear stress mechanotransduction (4).

Article Snippet: To assess the contribution of glypican-1 to mechanosensation, cells were transfected (Lipofectamine RNAiMAX, Invitrogen, 13778150) with an siRNA against glypican-1 (SR301881CL, OriGene) or a scrambled control (SR30004, OriGene) and exposed to static or shear stress (15 dynes/cm 2 ) conditions for 24 hr using the ibidi flow system described above.

Techniques: Activation Assay, Clinical Proteomics, Membrane, Binding Assay, Shear