Journal: Pediatric Research

Article Title: S100A12 on Circulating Endothelial Cells Surface in Children With Kawasaki Disease

doi: 10.1203/pdr.0b013e3181e67ce8

Figure Lengend Snippet: Figure 1. Diagram of CECs/MNC, CECs-S100A12/CECs, and FI- S100A12-CECs measurement by flow cytometry. A, screening MNC (R1); B, screening CECs (R2); C, verifying the CECs again (R3); D, measuring CECs-S100A12/CECs and FI-S100A12-CECs, the green represent the iso- type control scan.

Article Snippet: The samples were divided into two tubes (each with 100 L): one tube was used for testing by adding 10 L polyclonal goat anti-human S100A12 antibody (R&D systems), and the other was used as control by adding goat IgG1.

Techniques: Cytometry, Control

Journal: Pediatric Research

Article Title: S100A12 on Circulating Endothelial Cells Surface in Children With Kawasaki Disease

doi: 10.1203/pdr.0b013e3181e67ce8

Figure Lengend Snippet: Figure 2. Diagram of CECs/MNC, CECs-S100A12/CECs and FI-S100A12-CECs levels in Kawasaki disease with or without coronary artery lesions. A, CECs/MNC levels; B, CECs-S100A12/CECs levels; C, FI-S100A12-CECs levels. CECs/MNC, the ratio of circulating endothelial cells to MNC; CECs-S100A12/CECs, the positive rate of S100A12 on circulating endothelial cells surface; FI-S100A12-CECs, the fluorescence intensity of S100A12 on circulating endothelial cells surface; F, KD without CALs; f, KD with CALs; Œ, Control.

Article Snippet: The samples were divided into two tubes (each with 100 L): one tube was used for testing by adding 10 L polyclonal goat anti-human S100A12 antibody (R&D systems), and the other was used as control by adding goat IgG1.

Techniques: Control

Journal: Biochimica et biophysica acta

Article Title: RAGE overexpression confers a metastatic phenotype to the WM115 human primary melanoma cell line.

doi: 10.1016/j.bbadis.2014.02.013

Figure Lengend Snippet: Fig. 1. Q2 A) Levels of RAGE in RAGE transfected WM115 cells as determined by ELISA. Levels are expressed in pg RAGE protein per mg of total protein. WM115-RAGE and WM115-RAGE-I expressed 94 fold and 7 fold higher RAGE protein than the MOCK control cells, respectively. B) Binding of the anti-RAGE antibody MAB1145 to WM115-RAGE (filled circles) and WM115- MOCK (filled squares) measured by flow cytometry. The binding curve of MAB1145 to WM115-RAGE was fitted using a 1:1 binding model and showed an affinity of 1.5 (±0.3) nM. RAGE overexpressed in the melanoma cells is properly processed and translocated to the cell-surface, as demonstrated by their recognition by specific antibodies. The experiment was performed in triplicate and the standard deviation is indicated. C–F) Morphology of WM115-MOCK (C), WM115-RAGE-I (D), WM115-RAGE (E) and WM266-MOCK (F) by bright field microscopy. G–H) Differences in morphology between WM115-MOCK (G) and WM115-RAGE (H) transfected cells, as shown by actin staining. Actin was stained with PE conjugated phalloidin and the nuclei were stained with Hoechst 33342. (20× magnification).

Article Snippet: RAGE protein levels in the cell extracts were determined using the Quantikine human RAGE Immunoassay kit (R&D Systems) according to the manufacturer's procedure, and expressed in picogram of RAGE per mg of total protein.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay, Control, Binding Assay, Cytometry, Standard Deviation, Microscopy, Staining

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: Circulation Research

Article Title: S100A12 Mediates Aortic Wall Remodeling and Aortic Aneurysm

doi: 10.1161/circresaha.109.209486

Figure Lengend Snippet: Figure 5. S100A12 induces IL-6 and nuclear translocation of pSmad2. A, ELISA measuring IL-6 from cell culture supernatant from TG-VSMC and WT-VSMC cultures stimulated with 1 g/mL LPS and soluble RAGE as indicated. *P0.001 com- pared to baseline conditions; #P0.05 compared to LPS- stimulated condition. B, Immunoblotting with -S100A12 IgG using culture supernatant from TG-VSMCs (lanes 1 and 2) and WT-VSMCs (lanes 3 and 4) revealed S100A12 protein release into the cell culture supernatant only from TG VSMCs after treatment with LPS. C, IL-6 was also produced from aortic ring tissues (3 mm) after LPS exposure. *P0.03. D, IL-6 ELISA from mouse serum. *P0.05). E, Immunofluorescence microscopy of cultured primary VSMCs isolated from WT mice (top) and TG mice (middle and bottom) were stained with -pSmad2 IgG and DAPI. Scale bar20 m. Pretreatment with neutralizing TGF- (0.5 g/mL) is shown (bottom).

Article Snippet: Human full length S100A12 cDNA (Origene, Rockville, MD) was amplified by PCR using primers with added restriction site for Hind III (5’) and BamH I (3’) and ligated into the Bluescript II SK (Stratagene, La Jolla, CA) vector containing the smooth muscle cell promoter SM22.1, 2 Verification of the construct was obtained by restriction analysis and by sequencing.

Techniques: Translocation Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Produced, Microscopy, Isolation, Staining

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