Journal: iScience

Article Title: Host biomarker-based quantitative rapid tests for detection and treatment monitoring of tuberculosis and COVID-19.

doi: 10.1016/j.isci.2022.105873

Figure Lengend Snippet: Figure 1. Evaluation of host biomarkers for TB and LTBI in a European cohort Levels of IL-6, IP-10, ferritin, SAA1/A2, CRP, ApoA1, and S100A12 were measured by UCP-LFA in serum samples of TB patients (n = 30; green dots) and LTBI (n = 29; gray dots) from Europe. Median values for each group are indicated by horizontal bars. Mann-Whitney U tests were performed to determine the statistical significance between groups (pvalues: *p%0 $ 05, **p%0 $ 01, ***p%0 $ 001, ****p%0 $ 0001). Green dots: TB cohort 1; gray dots: LTBI cohort 1. AUC: area under the curve; Fc: flow control line; LTBI: latent tuberculosis infection; T: test line; TB: tuberculosis.

Article Snippet: 4 mm width UCP-LF strips specific for a single host protein – ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 - were produced as described earlier.24,28,29 For ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 LF strips, each Test (T) line comprised 200 ng of the following antibodies: goat-anti-human ApoA1 pAb (AF3664; R&D systems, Minneapolis, MN, USA), mouse-anti-human CRP mAb (C5; Labned.com, Amstelveen, the Netherlands), mouse-anti-human ferritin mAb (F31; Novus Biologicals, Littleton, CO, USA), rat-anti-human IL-6 mAb (MQ2-39C3; Biolegend, San Diego, CA, USA), mouse-anti-human IP-10 mAb (B-C55; Diaclone Research, Besancon, France), mouse-anti-human SAA1/A2 mAb (865504; R&D systems, Minneapolis, MN, USA), and goat-anti-human S100A12 pAb (AF1052; R&D systems, Minneapolis, MN, USA), respectively.

Techniques: MANN-WHITNEY, Control, Infection

Journal: iScience

Article Title: Host biomarker-based quantitative rapid tests for detection and treatment monitoring of tuberculosis and COVID-19.

doi: 10.1016/j.isci.2022.105873

Figure Lengend Snippet: Figure 2. Evaluation of host biomarkers for Dutch COVID-19 patients and healthy controls Levels of IL-6, IP-10, ferritin, SAA1/A2, CRP, ApoA1, and S100A12 were measured by UCP-LFA in serum samples of COVID-19 patients (n = 102) and healthy controls (n = 39; n = 27 from before (May) 2019 (n = 12 from after 2019 (June/July 2020)) from the Netherlands. Median values for each group are indicated by horizontal bars. Mann-Whitney U tests were performed to determine the statistical significance between groups (pvalues: *p%0 $ 05, **p%0 $ 01, ***p%0 $ 001, ****p %0 $ 0001). Orange dots: healthy controls from before 2019; purple dots: healthy controls from after 2019; black dots: COVID-19 patients with a fatal outcome; yellow dots: COVID-19 patients with severe disease; turquoise dots: COVID-19 patients with moderate disease. AUC: area under the curve; COVID-19: coronavirus disease 2019; Fc: flow control line; T: test line.

Article Snippet: 4 mm width UCP-LF strips specific for a single host protein – ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 - were produced as described earlier.24,28,29 For ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 LF strips, each Test (T) line comprised 200 ng of the following antibodies: goat-anti-human ApoA1 pAb (AF3664; R&D systems, Minneapolis, MN, USA), mouse-anti-human CRP mAb (C5; Labned.com, Amstelveen, the Netherlands), mouse-anti-human ferritin mAb (F31; Novus Biologicals, Littleton, CO, USA), rat-anti-human IL-6 mAb (MQ2-39C3; Biolegend, San Diego, CA, USA), mouse-anti-human IP-10 mAb (B-C55; Diaclone Research, Besancon, France), mouse-anti-human SAA1/A2 mAb (865504; R&D systems, Minneapolis, MN, USA), and goat-anti-human S100A12 pAb (AF1052; R&D systems, Minneapolis, MN, USA), respectively.

Techniques: MANN-WHITNEY, Control

Journal: iScience

Article Title: Host biomarker-based quantitative rapid tests for detection and treatment monitoring of tuberculosis and COVID-19.

doi: 10.1016/j.isci.2022.105873

Figure Lengend Snippet: Figure 3. Evaluation of host biomarkers for TB and COVID-19 patients Levels of IL-6, IP-10, ferritin, SAA1/A2, CRP, ApoA1, and S100A12 were measured by UCP-LFA in serum samples of TB patients (n = 46) and COVID-19 patients (n = 102) collected in European hospitals. Median values for each group are indicated by horizontal bars. Mann-Whitney U tests were performed to determine the statistical significance between groups (pvalues: *p%0 $ 05, **p%0 $ 01, ***p%0 $ 001, ****p%0 $ 0001). Green dots: TB cohort 1; blue dots: TB cohort 2; black dots: COVID-19 patients. AUC: area under the curve; COVID-19: coronavirus disease 2019; Fc: flow control line; T: test line; TB: tuberculosis.

Article Snippet: 4 mm width UCP-LF strips specific for a single host protein – ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 - were produced as described earlier.24,28,29 For ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 LF strips, each Test (T) line comprised 200 ng of the following antibodies: goat-anti-human ApoA1 pAb (AF3664; R&D systems, Minneapolis, MN, USA), mouse-anti-human CRP mAb (C5; Labned.com, Amstelveen, the Netherlands), mouse-anti-human ferritin mAb (F31; Novus Biologicals, Littleton, CO, USA), rat-anti-human IL-6 mAb (MQ2-39C3; Biolegend, San Diego, CA, USA), mouse-anti-human IP-10 mAb (B-C55; Diaclone Research, Besancon, France), mouse-anti-human SAA1/A2 mAb (865504; R&D systems, Minneapolis, MN, USA), and goat-anti-human S100A12 pAb (AF1052; R&D systems, Minneapolis, MN, USA), respectively.

Techniques: MANN-WHITNEY, Control

Journal: iScience

Article Title: Host biomarker-based quantitative rapid tests for detection and treatment monitoring of tuberculosis and COVID-19.

doi: 10.1016/j.isci.2022.105873

Figure Lengend Snippet: Figure 4. Treatment monitoring for TB Levels of IL-6, IP-10, ferritin, SAA1/A2, CRP, ApoA1, and S100A12 were measured by UCP-LFA in serum samples of pulmonary TB patients (n = 22) before treatment (t0), at months 2–4 (t1), and months 5–9 (t2) of treatment. Median values for each group are indicated by horizontal bars. The gray dotted lines represent the median value of the corresponding marker measured for 39 healthy controls. S100A12 data were missing for one patient. Friedman test with Dunn’s correction for multiple testing was performed to determine the statistical significance between timepoints (pvalues: *p%0 $ 05, **p%0 $ 01, ***p%0 $ 001, ****p%0 $ 0001). Fc: flow control line; T: test line; TB: tuberculosis; t0: first timepoints; t1: 2–4 months after the beginning of treatment; t2: 5–9 months after the beginning of treatment.

Article Snippet: 4 mm width UCP-LF strips specific for a single host protein – ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 - were produced as described earlier.24,28,29 For ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 LF strips, each Test (T) line comprised 200 ng of the following antibodies: goat-anti-human ApoA1 pAb (AF3664; R&D systems, Minneapolis, MN, USA), mouse-anti-human CRP mAb (C5; Labned.com, Amstelveen, the Netherlands), mouse-anti-human ferritin mAb (F31; Novus Biologicals, Littleton, CO, USA), rat-anti-human IL-6 mAb (MQ2-39C3; Biolegend, San Diego, CA, USA), mouse-anti-human IP-10 mAb (B-C55; Diaclone Research, Besancon, France), mouse-anti-human SAA1/A2 mAb (865504; R&D systems, Minneapolis, MN, USA), and goat-anti-human S100A12 pAb (AF1052; R&D systems, Minneapolis, MN, USA), respectively.

Techniques: Marker, Control

Journal: iScience

Article Title: Host biomarker-based quantitative rapid tests for detection and treatment monitoring of tuberculosis and COVID-19.

doi: 10.1016/j.isci.2022.105873

Figure Lengend Snippet: Figure 5. Treatment monitoring for COVID-19 Levels of IL-6, IP-10, ferritin, SAA1/A2, CRP, ApoA1, and S100A12 were measured by UCP-LFA in serum samples from COVID-19 patients (n = 25) at hospital admission (t0) and follow-up (t2). Median values for each group are indicated by horizontal bars. The gray dotted lines represent the median value of the corresponding marker measured for 39 healthy controls. Wilcoxon matched pairs signed rank tests were performed to determine the statistical significances between timepoints (pvalues: *p%0 $ 05, **p%0 $ 01, ***p%0 $ 001, ****p%0 $ 0001). COVID-19: coronavirus disease 2019; Fc: flow control line; T: test line; t0: timepoint of hospital admission; t2: follow-up around 6 weeks after hospital discharge.

Article Snippet: 4 mm width UCP-LF strips specific for a single host protein – ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 - were produced as described earlier.24,28,29 For ApoA1, CRP, ferritin, IL-6, IP-10, SAA1/A2, and S100A12 LF strips, each Test (T) line comprised 200 ng of the following antibodies: goat-anti-human ApoA1 pAb (AF3664; R&D systems, Minneapolis, MN, USA), mouse-anti-human CRP mAb (C5; Labned.com, Amstelveen, the Netherlands), mouse-anti-human ferritin mAb (F31; Novus Biologicals, Littleton, CO, USA), rat-anti-human IL-6 mAb (MQ2-39C3; Biolegend, San Diego, CA, USA), mouse-anti-human IP-10 mAb (B-C55; Diaclone Research, Besancon, France), mouse-anti-human SAA1/A2 mAb (865504; R&D systems, Minneapolis, MN, USA), and goat-anti-human S100A12 pAb (AF1052; R&D systems, Minneapolis, MN, USA), respectively.

Techniques: Marker, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Human Plasma Transcriptome Implicates Dysregulated S100A12 Expression: A Strong, Early-Stage Prognostic Factor in ST-Segment Elevated Myocardial Infarction: Bioinformatics Analysis and Experimental Verification

doi: 10.3389/fcvm.2022.874436

Figure Lengend Snippet: Ninety-one DEGs were identified from GSE60993 and GSE61144 microarrays for STEMI.

Article Snippet: The concentration of S100A12 was then determined using commercially available ELISA kits (Human S100A12 DuoSet ELISA kit, Catalog # DY1052-05; R&D Systems, Inc., Minneapolis, MN, United States) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Human Plasma Transcriptome Implicates Dysregulated S100A12 Expression: A Strong, Early-Stage Prognostic Factor in ST-Segment Elevated Myocardial Infarction: Bioinformatics Analysis and Experimental Verification

doi: 10.3389/fcvm.2022.874436

Figure Lengend Snippet: Univariate analysis of survival to discharge in our validated cohort.

Article Snippet: The concentration of S100A12 was then determined using commercially available ELISA kits (Human S100A12 DuoSet ELISA kit, Catalog # DY1052-05; R&D Systems, Inc., Minneapolis, MN, United States) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Human Plasma Transcriptome Implicates Dysregulated S100A12 Expression: A Strong, Early-Stage Prognostic Factor in ST-Segment Elevated Myocardial Infarction: Bioinformatics Analysis and Experimental Verification

doi: 10.3389/fcvm.2022.874436

Figure Lengend Snippet: A univariate and multivariate logistic regression model evaluating the association of clinical factors with in-hospital mortality in our validated cohort.

Article Snippet: The concentration of S100A12 was then determined using commercially available ELISA kits (Human S100A12 DuoSet ELISA kit, Catalog # DY1052-05; R&D Systems, Inc., Minneapolis, MN, United States) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Human Plasma Transcriptome Implicates Dysregulated S100A12 Expression: A Strong, Early-Stage Prognostic Factor in ST-Segment Elevated Myocardial Infarction: Bioinformatics Analysis and Experimental Verification

doi: 10.3389/fcvm.2022.874436

Figure Lengend Snippet: Clinical characteristics of patients with MACE during follow-up in our validated cohort.

Article Snippet: The concentration of S100A12 was then determined using commercially available ELISA kits (Human S100A12 DuoSet ELISA kit, Catalog # DY1052-05; R&D Systems, Inc., Minneapolis, MN, United States) according to the manufacturer’s instructions.

Techniques:

Journal: The FASEB Journal

Article Title: Structural and molecular characteristics of weight‐bearing volar skin can be reconstituted by micro skin tissue column grafting

doi: 10.1096/fj.202400866R

Figure Lengend Snippet: Distinctive structural and molecular characteristics of porcine volar skin. (A) Trichrome‐stained tissue sections from porcine trunk (left) and volar skin (right) are shown at the same magnification for comparison. Note the great difference in thickness between the epidermis in trunk skin (bracket #1) versus volar skin (brackets #3 and 4). The trunk stratum corneum is too thin to see in the main image and is therefore shown in the inset (bracket #2), displaying the classic layered structure. In contrast, the volar stratum corneum (bracket #4) is greatly thickened and has a compact structure. (B) IHC staining (red) showing differential expression of marker proteins in trunk versus volar (plantar) skin, as labeled in the figure. Porcine volar skin is distinguished by upregulation of S100A8, expression of S100A12, altered spatial distribution of S100A14 (strong pericellular staining in the viable epidermis with weak staining in the stratum corneum in plantar skin, vs. strong stratum corneum staining in trunk skin) and downregulation of STIM1. Insets show the plantar epidermis (specifically the stratum spinosum ) at higher magnification. Scale bars: A: 1 mm; inset, 25 μm; B: Trunk, 25 μm; plantar, 1 mm; insets, 50 μm.

Article Snippet: Immunohistochemical staining was performed as previously described, using antibodies against Keratin 7 (Abcam, ab68460, 1:100), Keratin 9 (Abcam, ab171966, 1:200), Vimentin (Vector Laboratories, VP‐RM17, 1:100), S100A8 (MyBiosource, MBS2028565, San Diego, CA, 1:400), S100A12 (Novus Biologicals, NBP1‐86694, Centennial, CO, 1:250), S100A14 (ThermoFisher, PA5‐55666, Waltham, MA, 1:2500), and STIM1 (Novus Biologicals, NBP110‐60547, Centennial, CO, 1:200).

Techniques: Staining, Comparison, Immunohistochemistry, Quantitative Proteomics, Marker, Labeling, Expressing

Journal: The FASEB Journal

Article Title: Structural and molecular characteristics of weight‐bearing volar skin can be reconstituted by micro skin tissue column grafting

doi: 10.1096/fj.202400866R

Figure Lengend Snippet: Volar characteristics recapitulated in ectopic graft sites. (A) Photographs of graft sites taken at various timepoints after injury and MSTC grafting, as denoted in the figure. Each photograph is shown at 2.5 mm across. The wound edges were tattooed with black ink to facilitate tracking of wound contraction. (B–E) Trichrome staining of graft sites at week 8. (B) Graft site treated with volar MSTCs, edges of graft site marked by arrowheads. (C) High‐power view of a volar MSTC‐treated graft site, focusing on the transition zone between the volar‐treated wound area (left of the dotted line) and the neighboring skin (right of the dotted line). Note substantially thickened epidermis, structurally compact stratum corneum, as well as pronounced rete ridges (arrows). (D) Control wound site histology, with edges of the injured area marked by arrowheads. (E) High‐power view of control wound, showing the transition zone between area of injury (left of the dotted line) and the neighboring skin (right of the dotted line). Note the epidermis and stratum corneum are similar in both thickness and structure between the injured area and surrounding skin, as well as the paucity of rete ridges in the area of injury. (F) Wound area measured at week 8, showing less contraction in the MSTC‐treated group compared to control. (G) The ratio between the lengths of the dermal‐epidermal junction (DEJ) and the top of the viable epidermis was taken to reflect the extent of rete ridges, which was about 2.5x greater in the volar MSTC‐treated wounds. (H) The stratum corneum was also significantly thicker in the volar MSTC‐treated wounds than in controls. (I–L) IHC staining of volar MSTC‐treated sites for various volar markers, as indicated in the images. Images are focused on the transition zone between the volar‐treated wound area (left of the dotted line) and the neighboring skin (right of the dotted line). (I) Strong S100A8 expression, most notably in the stratum corneum, in the volar MSTC‐treated wound area, but not in neighboring skin. (J) Expression of S100A12 only in the volar MSTC‐treated wound area, but not in neighboring skin. (K) The volar‐treated wound area shows strong pericellular S100A14 expression in the viable epidermis but weak expression in the stratum corneum, as opposed to the neighboring skin, where S100A14 expression is much stronger in the stratum corneum. (L) STIM1 expression is reduced in the volar MSTC‐treated wound area. (M–R) Quantification of IHC staining intensity for the different markers of interest. (M–P) show results from the viable epidermis. (Q and R) show results in the stratum corneum. In summary, the structural and molecular changes are consistent with characteristics of volar skin, and are highly specific to the volar MSTC‐treated areas, indicating that they are not a mere byproduct of the general wound healing response. * p < .05; ** p < .01; *** p = .0001; **** p < .0001; ns, not significant. Scale bars: B: 1 mm; C: 250 μm; D: 1 mm; I–L: 250 μm.

Article Snippet: Immunohistochemical staining was performed as previously described, using antibodies against Keratin 7 (Abcam, ab68460, 1:100), Keratin 9 (Abcam, ab171966, 1:200), Vimentin (Vector Laboratories, VP‐RM17, 1:100), S100A8 (MyBiosource, MBS2028565, San Diego, CA, 1:400), S100A12 (Novus Biologicals, NBP1‐86694, Centennial, CO, 1:250), S100A14 (ThermoFisher, PA5‐55666, Waltham, MA, 1:2500), and STIM1 (Novus Biologicals, NBP110‐60547, Centennial, CO, 1:200).

Techniques: Staining, Control, Immunohistochemistry, Expressing

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: 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: Current issues in molecular biology

Article Title: Increased Production of Interleukin-10 and Tumor Necrosis Factor-Alpha in Stimulated Peripheral Blood Mononuclear Cells after Inhibition of S100A12.

doi: 10.3390/cimb44040117

Figure Lengend Snippet: Figure 1. Plasma levels of advanced glycation endproducts (AGE), soluble RAGE (sRAGE), high- mobility group box 1 (HMGB1), and S100A12 on Days 1 and 7 are shown using bar charts with one standard error. In survivors, plasma AGE levels were significantly increased and plasma sRAGE levels were significantly decreased after 6 d. There were no changes in plasma levels of AGE and sRAGE in non-survivors. Plasma levels of HMGB1 and S100A12 did not change in survivors and non-survivors after 6 d.

Article Snippet: Cells in the 3rd well were stimulated with 1 μg/mL LPS (Sigma, Burlington, MO, USA) and treated with 250 pg/mL neutralizing anti-S100A12 human antibody (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Clinical Proteomics

Journal: Current issues in molecular biology

Article Title: Increased Production of Interleukin-10 and Tumor Necrosis Factor-Alpha in Stimulated Peripheral Blood Mononuclear Cells after Inhibition of S100A12.

doi: 10.3390/cimb44040117

Figure Lengend Snippet: Figure 2. Bar charts with one standard error show supernatant levels of tumor necrosis factor (TNF)- α, interleukin (IL)-10, and IL-12 from peripheral blood mononuclear cells (PBMCs) with and without lipopolysaccharide (LPS) stimulation and anti-S100A12 treatment. Productions of TNF-α, IL-10, and IL-12 from PBMCs were significantly increased after LPS stimulation in both controls and patients with sepsis. Additional anti-S100A12 monoclonal antibodies increased the production of TNF-α and IL-10 in LPS-stimulated PBMCs from both controls and patients.

Article Snippet: Cells in the 3rd well were stimulated with 1 μg/mL LPS (Sigma, Burlington, MO, USA) and treated with 250 pg/mL neutralizing anti-S100A12 human antibody (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Bioprocessing

Journal: Current issues in molecular biology

Article Title: Increased Production of Interleukin-10 and Tumor Necrosis Factor-Alpha in Stimulated Peripheral Blood Mononuclear Cells after Inhibition of S100A12.

doi: 10.3390/cimb44040117

Figure Lengend Snippet: Figure 3. A proposed schematic mechanism that shows S100A12 may modulate tumor necrosis factor (TNF)-α and interleukin (IL)-10 production through lipopolysaccharide (LPS). LPS links Toll- like receptor (TLR) 4 to activate nuclear factor-kappa B (NF-κB) through myeloid differentiation primary response 88 (MYD88). The activated NF-κB in the cytoplasm is then translocated into the nucleus where it binds to specific sequences of DNA and increases IL-10, IL-12, and TNF-α gene expressions. Extracellular IL-10 binds to IL-10 receptors and activates signal transducer and activator of transcription (STAT)3. Then, STAT3 is translocated to the cell nucleus and induces IL-10 gene expression. Activating STAT3 signaling also induces suppressor of cytokine signaling (SOCS)3 to suppress IL-12 and TNF-α gene expression. Once RAGE is bound to LPS or S100A12, NF-κB is activated by the active form of rat sarcoma (Ras), Ras nucleotide guanosine triphosphate (GTP) hydrolases (GTPase). S100A12 may competitively bind to TLR4/RAGE and the affinity between S100A12 and TLR4/RAGE may be lower than that between LPS and TLR/4RAGE. This results in increased IL-10 and TNF-α production with S100A12 being inhibited.

Article Snippet: Cells in the 3rd well were stimulated with 1 μg/mL LPS (Sigma, Burlington, MO, USA) and treated with 250 pg/mL neutralizing anti-S100A12 human antibody (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Gene Expression