Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Functional Role of Soluble Receptor for Advanced Glycation End Products in Stroke

doi: 10.1161/atvbaha.112.300523

Figure Lengend Snippet: Figure 1. Plasma levels of soluble form of receptors for advanced glycation end products (sRAGE) and high mobility group box 1 (HMGB1) in acute ischemic stroke (IS) patients. Plasma levels of sRAGE increase significantly at 48 hours after stroke, compared with controls, and then decrease at 5 to 7 days (A). Plasma levels of HMGB1 are significantly higher in IS patients than in controls at all 3 time points (B). #P<0.05 vs control. *P<0.05 vs <48-hour samples.

Article Snippet: Mice were administered 1 mg/kg of sRAGE (1179RG, R&D Systems, Inc), 1 mg/kg of HMGB1 (Hmgb1-2628 mol/L, Creative BioMart), 1 mg/kg of sRAGE + 1 mg/kg of HMGB1, or vehicle (saline) by infusion into the femoral vein (100 μL) 180 minutes after the start of reperfusion.

Techniques: Clinical Proteomics, Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Functional Role of Soluble Receptor for Advanced Glycation End Products in Stroke

doi: 10.1161/atvbaha.112.300523

Figure Lengend Snippet: Figure 2. Expression of receptor for advanced glycation end products (RAGE) and high mobility group box 1 (HMGB1) after stroke. Membrane RAGE and HMGB1 levels increase markedly as early as 3 hours and even higher at 24 and 72 hours in ipsilateral brain tissues after ischemia and reperfusion (I/R) vs sham controls (A through C). Plasma expression of soluble form of RAGE (sRAGE) significantly increases at 24 hours after I/R (D and E) and plasma levels of HMGB1 at all 3 time points after I/R (D and F). Immunoprecipitation shows the amount of plasma HMGB1 binding to sRAGE increases after stroke both in mice (G) and IS patients (H). *P<0.05 vs sham levels. Sta- tistical comparisons were made with ANOVA, followed by Newman–Keuls post hoc analysis.

Article Snippet: Mice were administered 1 mg/kg of sRAGE (1179RG, R&D Systems, Inc), 1 mg/kg of HMGB1 (Hmgb1-2628 mol/L, Creative BioMart), 1 mg/kg of sRAGE + 1 mg/kg of HMGB1, or vehicle (saline) by infusion into the femoral vein (100 μL) 180 minutes after the start of reperfusion.

Techniques: Expressing, Membrane, Clinical Proteomics, Immunoprecipitation, Binding Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Functional Role of Soluble Receptor for Advanced Glycation End Products in Stroke

doi: 10.1161/atvbaha.112.300523

Figure Lengend Snippet: Figure 4. In vitro study shows exogenous soluble form of receptors for advanced glycation end products (RAGE; sRAGE; 50 ng/mL) significantly protects primary cortical neurons from glucose deprivation (GD) and oxygen GD (OGD) injury, and premixing sRAGE with high mobility group box 1 (HMGB1) can reverse the detrimental effect of HMGB1 (A and B). **P<0.05 vs Locke buffer control cultures. RAGE immunoreactivities in cultured neurons in normal (Neural basal) or following GD or OGD plus reperfusion conditions; cells were counterstained with 4'-6-diamidino-2-phenylindole to label all nuclei and with the neuron-specific marker mitogen-activated protein (MAP) 2 (C). Exogenous sRAGE in culture medium significantly reduces activation of c-Jun amino-terminal kinase (JNK; D and E), consequent caspase 3 activation (D and F), and nuclear translocation of p65 protein (G and H) in cortical neurons following 6- and 24-hour GD condi- tion vs controls. Premixing sRAGE with HMGB1 ameliorates the aggravating effect of HMGB1 on JNK and nuclear factor-κB activities in neurons subjected to GD (D through H). *P<0.05 vs Locke’s buffer control cultures. **P<0.01 vs Locke buffer control cultures. Statistical comparisons were made with ANOVA, followed by Newman–Keuls post hoc analysis.

Article Snippet: Mice were administered 1 mg/kg of sRAGE (1179RG, R&D Systems, Inc), 1 mg/kg of HMGB1 (Hmgb1-2628 mol/L, Creative BioMart), 1 mg/kg of sRAGE + 1 mg/kg of HMGB1, or vehicle (saline) by infusion into the femoral vein (100 μL) 180 minutes after the start of reperfusion.

Techniques: In Vitro, Control, Cell Culture, Marker, Activation Assay, Translocation Assay

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: SC79 induces the shedding of the RAGE ectodomain. HAECs were incubated with 10 µM SC79 for various times (5, 10, 30, and 60 min) ( n = 4) ( A ) or different concentrations of SC79 (0.1, 1, 5, and 10 µM) for 30 min ( n = 3) ( B ). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and an anti-actin antibody. To compare the size of RAGE in cell lysate and culture supernatant, untreated cell lysate (a) and conditioned media from cells treated with 10 µM SC79 for 30 min (b) were run on the same gel and immunoblotted with the RAGE antibody ( C ). The cell lysates of HAECs treated with different concentrations of SC79 (0.1, 1, 5, and 10 µM) for 30 min were immunoblotted with an antibody to the C-terminal domain of human RAGE and an anti-actin antibody (n = 4) ( D ). ( * p < 0.05 vs. control)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Incubation, Control

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: Inhibitors of AKT and ADAM10 diminish SC79-induced RAGE ectodomain shedding. HAECs were preincubated with or without MK-2206 (1 µM), GI 254023X (2 µM), or DMSO (vehicle) for 60 min. Following this, they were further incubated for 30 min with or without SC79 (10 µM). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and an anti-actin antibody. ( n = 3, * p < 0.05 vs. control, # p < 0.05 vs. SC79 treatment alone)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Incubation, Control

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: AKT1 activation is required for SC79-induced RAGE ectodomain shedding. ( A ) HAECs express all three AKT isoforms, and AKT1-, AKT2-, and AKT3-siRNAs selectively deplete each AKT isoform. HAECs were transfected with AKT1-, AKT2-, AKT3-siRNAs, or control siRNAs, and the cell lysates were immunoblotted with antibodies to AKT1, AKT2, AKT3, or actin. ( n = 3, * p < 0.05 vs. control). ( B ) SC79 activates AKT1. HAECs were incubated with 10 µM SC79 for various times (1, 5, 10, and 30 min) (upper panel) or different concentrations of SC79 (0.1, 1, 5, and 10 µM) for 30 min (lower panel). The cell lysates were immunoblotted with antibodies to p-AKT1 (Ser473) and AKT1. ( n = 3, * p < 0.05 vs. control). ( C ) AKT1 knockdown inhibits SC79-induced RAGE ectodomain shedding. HAECs were transfected with AKT1-siRNA or control siRNA and then incubated for 30 min with or without SC79 (10 µM). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and antibodies to AKT1 and actin. ( n = 3, * p < 0.05 vs. control cells transfected with control siRNA). ( D ) AKT1 knockdown abolishes SC79’s inhibitory effect against AGE-BSA. HAECs transfected with AKT1-siRNA or control siRNA were treated for 30 min with or without SC79 (10 µM). The cells were then treated with AGE-BSA (100 µg/ml) for 24 h. The cell lysates were immunoblotted with antibodies to ICAM-1, AKT1, and actin. ( n = 3, * p < 0.05 vs. control; # p < 0.05 vs. AGE-BSA; † p < 0.05 vs. control cells transfected with AKT1-siRNA)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Activation Assay, Transfection, Control, Incubation, Knockdown

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: AKT2 activation is required for SC79-induced RAGE ectodomain shedding. ( A ) SC79 activates AKT2. HAECs were incubated with 10 µM SC79 for various times (1, 5, 10, and 30 min) (upper panel) or different concentrations of SC79 (0.1, 1, 5, and 10 µM) for 30 min (lower panel). The cell lysates were immunoblotted with antibodies to p-AKT2 (Ser474) and AKT2. ( n = 4, * p < 0.05 vs. control). ( B ) AKT2 knockdown inhibits SC79-induced RAGE ectodomain shedding. HAECs were transfected with AKT2-siRNA or control siRNA and then incubated for 30 min with or without SC79 (10 µM). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and antibodies to AKT2 and actin. ( n = 4, * p < 0.05 vs. control cells transfected with control siRNA). ( C ) AKT2 knockdown abolishes SC79’s inhibitory effect against AGE-BSA. HAECs transfected with AKT2-siRNA or control siRNA were treated for 30 min with or without SC79 (10 µM). The cells were then treated with AGE-BSA (100 µg/ml) for 24 h. The cell lysates were immunoblotted with antibodies to ICAM-1, AKT2, and actin. ( n = 3, * p < 0.05 vs. control; # p < 0.05 vs. AGE-BSA; † p < 0.05 vs. control cells transfected with AKT2-siRNA)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Activation Assay, Incubation, Control, Knockdown, Transfection

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: AKT3 activation is required for SC79-induced RAGE ectodomain shedding. ( A ) SC79 activates AKT3. HAECs were incubated with 10 µM SC79 for various times (1, 5, 10, and 30 min) (upper panel) or different concentrations of SC79 (0.1, 1, 5, and 10 µM) for 30 min (lower panel). The cell lysates were immunoblotted with antibodies to p-AKT3 (Ser472) and AKT3. ( n = 3, * p < 0.05 vs. control). ( B ) AKT3 knockdown inhibits SC79-induced RAGE ectodomain shedding. HAECs were transfected with AKT3-siRNA or control siRNA and then incubated for 30 min with or without SC79 (10 µM). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and antibodies to AKT3 and actin. ( n = 3, * p < 0.05 vs. control cells transfected with control siRNA). ( C ) AKT3 knockdown abolishes SC79’s inhibitory effect against AGE-BSA. HAECs transfected with AKT3-siRNA or control siRNA were treated for 30 min with or without SC79 (10 µM). The cells were then treated with AGE-BSA (100 µg/ml) for 24 h. The cell lysates were immunoblotted with antibodies to ICAM-1, AKT3, and actin. ( n = 3, * p < 0.05 vs. control; # p < 0.05 vs. AGE-BSA; † p < 0.05 vs. control cells transfected with AKT3-siRNA)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Activation Assay, Incubation, Control, Knockdown, Transfection

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: SC79 induces RAGE ectodomain shedding by promoting ADAM10 cell surface translocation. ( A ) Immunofluorescence staining to evaluate the effect of SC79 on ADAM10 localization. HAECs grown in culture dishes with a coverslip were treated with SC79 (10 µM) for 10–120 min. (a) The cells on the coverslip were fixed for 10 min with 4% paraformaldehyde without permeabilization, then immunostained with an antibody to an extracellular portion of ADAM10 and examined using confocal microscopy. DAPI was used to label the nuclei of the cells. Representative photos and the relative fluorescence intensities are shown (scale bar: 100 μm). (b) Cell lysates from cells that were not on the coverslip in the same culture plate were immunoblotted with antibodies to ADAM10 and actin. ( n = 3, * p < 0.05 vs. control). ( B ) ADAM10 knockdown inhibits SC79-induced RAGE ectodomain shedding. HAECs were transfected with ADAM10-siRNA or control siRNA and then incubated for 30 min with or without SC79 (10 µM). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and antibodies to ADAM10 and actin. ( n = 3, * p < 0.05 vs. control cells transfected with control siRNA). ( C ) ADAM10 knockdown abolishes SC79’s inhibitory effect against AGE-BSA. HAECs transfected with ADAM10-siRNA or control siRNA were treated for 30 min with or without SC79 (10 µM). The cells were then treated with AGE-BSA (100 µg/ml) for 24 h. The cell lysates were immunoblotted with antibodies to ICAM-1, ADAM10, and actin. ( n = 3, * p < 0.05 vs. control; # p < 0.05 vs. AGE-BSA; † p < 0.05 vs. control cells transfected with ADAM10-siRNA)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Translocation Assay, Immunofluorescence, Staining, Confocal Microscopy, Fluorescence, Control, Knockdown, Transfection, Incubation

Journal: Scientific Reports

Article Title: AKT activation triggers Rab14-mediated ADAM10 translocation to the cell surface in human aortic endothelial cells

doi: 10.1038/s41598-025-90624-w

Figure Lengend Snippet: Rab14 is required for SC79-induced ADAM10 cell surface translocation. ( A ) Rab14 knockdown prevents SC79-induced ADAM10 cell surface translocation. HAECs grown in culture dishes with a coverslip were transfected with Rab14-siRNA or control siRNA and then incubated for 20 min with DMSO or SC79 (10 µM). (a) Cells grown on the coverslip were immunostained with an antibody to an extracellular portion of ADAM10. Representative photos and the relative fluorescence intensities are shown (scale bar: 100 μm). (b) Cell lysates from cells that were not on the coverslip in the same culture plate were immunoblotted with antibodies to Rab14 and actin. ( n = 3, * p < 0.05 vs. control cells transfected with control siRNA). ( B ) Rab14 knockdown inhibits SC79-induced RAGE ectodomain shedding. HAECs were transfected with Rab14-siRNA or control siRNA and then incubated for 30 min with or without SC79 (10 µM). The cell lysate and culture supernatant were immunoblotted with a monoclonal antibody to the extracellular domain of human RAGE and antibodies to Rab14 and actin. ( n = 3, * p < 0.05 vs. control cells transfected with control siRNA). ( C ) Rab14 knockdown abolishes SC79’s inhibitory effect against AGE-BSA. HAECs transfected with Rab14-siRNA or control siRNA were treated for 30 min with or without SC79 (10 µM). The cells were then treated with AGE-BSA (100 µg/ml) for 24 h. The cell lysates were immunoblotted with antibodies to ICAM-1, Rab14, and actin. ( n = 4, * p < 0.05 vs. control; # p < 0.05 vs. AGE-BSA; † p < 0.05 vs. control cells transfected with Rab14-siRNA)

Article Snippet: Anti-RAGE (JF0975) rabbit monoclonal antibody against amino acids 350–390 corresponding to the C-terminal region of human RAGE was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Translocation Assay, Knockdown, Transfection, Control, Incubation, Fluorescence

Journal: Molecular metabolism

Article Title: Dietary sugars, not lipids, drive hypothalamic inflammation.

doi: 10.1016/j.molmet.2017.06.008

Figure Lengend Snippet: Figure 3: RAGE and ALCAM are expressed on non-neuronal cell populations. (A) CML binds to proteins of RAGE (molecular weight z 75 KDa) and ALCAM (molecular weight z 105 KDa); the vehicle does not bind to protein of either receptor, as illustrated by no band detected in binding of ALCAM. (B) RAGE and ALCAM gene expression in mediobasal hypothalami of chow or HCHF mice (n ¼ 6 for chow or for HCHF, P ¼ 0.019 for RAGE, P ¼ 0.006 for ALCAM). (C) RAGE is intensely expressed by microglia (iba1-ir, indicated by white arrowheads). Higher magnifications of the areas framed by dashed lines are presented in D. (E) RAGE is intensely expressed on endothelial cells (laminin-ir, indicated by white arrows). Higher magnifications of the areas framed by dashed lines are presented in F. (G) CML stimulates TNFa, but not PDGF-B, gene expression in cultured primary microglia (n ¼ 6 wells of cells for vehicle, n ¼ 5 for TNFa treatments, P ¼ 0.04 for TNFa). (H & I) CML stimulates microglial reactivity in the mediobasal hypothalamic area, arrowheads point to the areas where the tip of the infusion probes located. (J) Iba1-ir cell number and cell coverage in H & I (n ¼ 4 mice for vehicle, n ¼ 5 for CML). (K) ALCAM is expressed on part of the vasculature (laminin-ir, indicated by white arrows, two pericytes are indicated by white arrowheads); higher magnifications of the areas framed by dashed lines are presented in L. (M) ALCAM is expressed on pericytes (PDGFRb-ir, white arrowheads). Higher magnifications of the areas framed by dashed lines are presented in N. Scale bar: 30 mm in C, E, K and M, 7.5um in D, F, L and N. Data are presented as means s.e.m. *P < 0.05, **P < 0.01. P values for unpaired comparisons were analyzed by two-tailed Student’s t test.

Article Snippet: Briefly, 2 mg recombinant CD166/ALCAM protein (Recombinant Mouse ALCAM/CD166 Fc Chimera; R&D Systems) and RAGE protein (Recombinant Mouse RAGE Fc Chimera R&D Systems) were separated on 10% SDS gels (Bio-rad cat.567-1033) and transferred to PVDF membranes.

Techniques: Molecular Weight, Binding Assay, Gene Expression, Cell Culture, Two Tailed Test

Journal: Molecular metabolism

Article Title: Dietary sugars, not lipids, drive hypothalamic inflammation.

doi: 10.1016/j.molmet.2017.06.008

Figure Lengend Snippet: Figure 4: Deletions of RAGE or ALCAM genes improve metabolic symptoms induced by a HCHF diet and exert diverse impacts on microglia, pericytes, and vasculature in the arcuate nucleus. (A & B) Daily caloric intake (in wk10) and weekly BW gain of chow or HCHF diet-fed WT versus RAGE/ mice (n ¼ 5e8 per group); For weekly BW gain, in all time points, WT and RAGE/ mice have less BW gain on chow diet than on HCHF diet (P < 0.0001); from wk14 to wk16, BW gain on HCHF of RAGE/ mice is significantly less than WT mice. (C & D) Daily caloric intake (in wk10) and weekly BW gain of chow or HCHF diet fed WT mice versus ALCAM/ mice (n ¼ 5e8 per group). For weekly BW gain, from wk2 on, WT and ALCAM/ mice have less BW gain in chow than in HCHF; from wk12, wk14 to wk16, BW-gain on HCHF of ALCAM/ mice is significantly less than WT mice. (E & F) Quantification of the number of iba1-ir microglia and the PDGFRb-ir pericytes in the ARC in chow or HCHF diet fed WT mice versus RAGE/ mice (n ¼ 5e9 per group). (G & H) Quantification of vessel length and vascular density in the ARC from chow or HCHF diet fed WT mice versus RAGE/ mice (n ¼ 5e7 per group). (I & J) Quantification of the number of iba1-ir microglia and the PDGFRb-ir pericytes in the ARC in chow or HCHF diet fed WT mice versus ALCAM/ mice (n ¼ 5e6 per group). (K & L) Quantification of vessel length and vascular density in the ARC from chow or HCHF diet fed WT mice versus ALCAM/ mice (n ¼ 6 per group). Data are presented as means s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. Asterisks in B and D indicate significance between WT and RAGE/ or ALCAM/ mice on HCHF diet. Two-way ANOVA followed by Bonferroni multiple comparisons for post-hoc analysis was performed to detect significant interaction between genotype and diet on each parameter.

Article Snippet: Briefly, 2 mg recombinant CD166/ALCAM protein (Recombinant Mouse ALCAM/CD166 Fc Chimera; R&D Systems) and RAGE protein (Recombinant Mouse RAGE Fc Chimera R&D Systems) were separated on 10% SDS gels (Bio-rad cat.567-1033) and transferred to PVDF membranes.

Techniques:

Journal: Molecular metabolism

Article Title: Dietary sugars, not lipids, drive hypothalamic inflammation.

doi: 10.1016/j.molmet.2017.06.008

Figure Lengend Snippet: Figure 5: Deletion of RAGE and ALCAM genes improves metabolic symptoms induced by a HCHF diet. (A & B) Daily caloric intake (in wk10) and weekly body weight gain of chow or HCHF diet-fed WT versus RAGE-ALCAM/ mice (n ¼ 6e11 per group); for weekly BW gain, from wk5 on, WT and RAGE-ALCAM/ mice have less BW gain in chow than in HCHF; From wk1 to wk4 and from wk9 to wk16, there are significant effect of genotype on BW gain on HCHF. (C) Body composition of WT versus RAGE-ALCAM/ mice fed HCHF diet (n ¼ 4e8 per group). (D) Glucose tolerance of WT versus RAGE-ALCAM/ mice fed chow or HCHF diet (n ¼ 5e7 per group). (E) Glucose tolerance of RAGE-ALCAM/

Article Snippet: Briefly, 2 mg recombinant CD166/ALCAM protein (Recombinant Mouse ALCAM/CD166 Fc Chimera; R&D Systems) and RAGE protein (Recombinant Mouse RAGE Fc Chimera R&D Systems) were separated on 10% SDS gels (Bio-rad cat.567-1033) and transferred to PVDF membranes.

Techniques:

Journal: Molecular metabolism

Article Title: Dietary sugars, not lipids, drive hypothalamic inflammation.

doi: 10.1016/j.molmet.2017.06.008

Figure Lengend Snippet: Figure 6: Deletion of RAGE and ALCAM genes reduces microglial reactivity and neovasculature formation in the arcuate nucleus (A, B & C) Quantification of iba1-ir microglial number, coverage and the PDGFRb-ir pericytes number in the ARC from chow or HCHF diet fed WT (n ¼ 5e8 per group) versus RAGE-ALCAM/ mice (n ¼ 7e 10 per group). (D & E) Quantification of vessel length and vascular density in the ARC from chow or HCHF diet fed WT (n ¼ 5e7 per group) versus RAGE-ALCAM/ mice (n ¼ 5e 7 per group). (FeH) Illustrations of the iba1-ir microglia, PDGFRb-ir pericytes and FITC-albumin labeled vessels in WT versus RAGE-ALCAM/ mice fed chow or HCHF diet, with a frame of 0.2 mm 0.2 mm for quantifications in the ARC. (I) Illustration of the skeletonization of vessel in H for vascular density analysis. III: third cerebral ventricle. Scale bar: 50um in F and G, 100um in F. Data are presented as means s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA followed by Bonferroni multiple comparisons for post-hoc analysis was performed to detect significant interaction between genotype and diet on each parameter.

Article Snippet: Briefly, 2 mg recombinant CD166/ALCAM protein (Recombinant Mouse ALCAM/CD166 Fc Chimera; R&D Systems) and RAGE protein (Recombinant Mouse RAGE Fc Chimera R&D Systems) were separated on 10% SDS gels (Bio-rad cat.567-1033) and transferred to PVDF membranes.

Techniques: Labeling

Journal: Disease markers

Article Title: Fecal S100A12 in Healthy Infants and Children

doi: 10.1155/2013/873582

Figure Lengend Snippet: Faecal S100A12 concentrations in 56 healthy infants and children. Serial stools collected from the first day of life (meconium) to 6 months of age from 7 healthy infants (Population 1) and single stools collected from 49 children (Population 2) were utilised to measure faecal S100A12 concentrations by immunoassay.

Article Snippet: Dilutions of recombinant S100A12 (Recombinant Human EN-RAGE/S100A12, R&D Systems, Minneapolis, MN, USA) were added (100 μ L/well) in order to create a standard curve.

Techniques:

Journal: Disease markers

Article Title: Fecal S100A12 in Healthy Infants and Children

doi: 10.1155/2013/873582

Figure Lengend Snippet: Measurement of fecal S100A12 infants and children. Repeated fecal samples were collected from seven term infants over the first six months of life (a). Single stool samples were collected from 49 healthy infants and children (b). S100A12 concentrations were measured by immunoassay. Only five samples (all in infants) were above the cut-off of 10 mg/kg.

Article Snippet: Dilutions of recombinant S100A12 (Recombinant Human EN-RAGE/S100A12, R&D Systems, Minneapolis, MN, USA) were added (100 μ L/well) in order to create a standard curve.

Techniques:

Journal: Molecules and Cells

Article Title: Characterization of αX I-Domain Binding to Receptors for Advanced Glycation End Products (RAGE)

doi: 10.14348/molcells.2017.0021

Figure Lengend Snippet: Binding of αX and αM I-domains to RAGE and the V-domain of RAGE. (A) A schematic representation of recombinant RAGE and RAGE derived soluble domains. All soluble proteins are fused with a His-tag for purification and detection. (B) SDS-PAGE analysis of purified sRAGE, sRAGEC1/2 and sRAGEV. (C) SPR sensorgram of sRAGE and RAGE-derived soluble domains binding to immobilized GST-αX-I. RAGE-derived proteins (1 μM) were injected to flow over immobilized GST-αX-I on a CM5 sensor chip (1800 RU). (D) Binding of sRAGEV and sRAGEC1/2 to GST-αX-I on microtiter plates. sRAGEV and sRAGEC1/2 (0.5 μM or 1.0 μM) were loaded on microtiter plates coated with GST-αX-I. Data are means ± S. E. (n = 3). (E, F) Binding of the I-domains to the sRAGE (E) and sRAGEV (F) on microtiter plates. GST and αX and αM I-domains (0.5 μM or 1.0 μM) were loaded on microtiter plates coated with sRAGE and sRAGEV. Data are means ± S. E. (n = 3).

Article Snippet: Recombinant RAGE fused with the human IgG Fc region produced from mammalian cells was purchased from R&D Systems (USA).

Techniques: Binding Assay, Recombinant, Derivative Assay, Purification, SDS Page, Injection

Journal: Experimental and Therapeutic Medicine

Article Title: Epigallocatechin-3-gallate attenuates neointimal hyperplasia in a rat model of carotid artery injury by inhibition of high mobility group box 1 expression

doi: 10.3892/etm.2017.4774

Figure Lengend Snippet: EGCG inhibits balloon injury-induced HMGB1 and RAGE expression levels. mRNA expression levels of (A) HMGB1 and (B) RAGE in artery tissues were determined by reverse transcription-quantitative polymerase chain reaction. Protein expression levels of (C) HMGB1 and (D) RAGE in artery tissues were detected by western blotting. β-actin was used as a loading control. The protein bands were quantified by gray scanning. Data are presented as the mean + standard deviation (n=6). *P<0.05, **P<0.01 and ***P<0.001 vs. the sham group; # P<0.05, ## P<0.01 and ### P<0.001 vs. the injury group. EGCG, epigallocatechin-3-gallate; HMGB1, high mobility group box 1; RAGE, receptor of advanced glycation end products.

Article Snippet: Subsequent to blocking with 5% skimmed milk for 1 h at room temperature, the membranes were incubated with primary antibodies against HMGB1 (Boster Biological Technology, Ltd., Wuhan, China; catalogue no. BA4277; 1:400), RAGE (Boster Biological Technology, Ltd.; catalogue no. PB0530; 1:400), nuclear factor (NF)-κB (Boster Biological Technology, Ltd.; catalogue no. BA0610; 1:400) and β-actin (Santa Cruz Biotechnology, Dallas, USA; catalogue no. sc-47778; 1:400), respectively at 4°C overnight.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control, Standard Deviation

Journal: Journal of Functional Foods

Article Title: Protective effects of selenium-enriched peptides from Cardamine violifolia on d-galactose-induced brain aging by alleviating oxidative stress, neuroinflammation, and neuron apoptosis

doi: 10.1016/j.jff.2020.104277

Figure Lengend Snippet: Fig. 7. CSP supplementation reduced D-gal-induced overexpression of RAGE, BACE-1, Aβ-42 and PS1. (A) Western blot analysis of RAGE, BACE-1, Aβ-42 and PS1; (B) The relative protein expressions of RAGE, BACE-1, Aβ-42 and PS1; Data are expressed as the mean ± SEM (n = 3). Differences were denoted as follows: * P < 0.05, ** P < 0.01, *** P < 0.001 compared with NC; # P < 0.05, ## p < 0.01, ### p < 0.001 compared with D-gal; & P < 0.05, && p < 0.01, &&& p < 0.001 compared with CSPL.

Article Snippet: Primary antibodies against NFkβ-p65, RAGE, BACE1, PS1, BAX, BCL2, Caspase-3, HO1, NQO1, and β-actin were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Over Expression, Western Blot