Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: Increased frequency and proinflammatory signature of CD48 high S100A12 + macrophages in rheumatoid arthritis synovium. A UMAP visualization of synovial macrophages clustered into nine subpopulations on the basis of scRNA-seq data. B Disease-stratified analysis showing an increased proportion of CD48 high S100A12 + macrophages in the RA synovium compared with those in the UA, OA, and HC groups. C UMAP plots depicting S100A12 expression intensity across macrophage subclusters in different disease states. D Gene Ontology (GO) enrichment of biological processes in CD48 high S100A12 + marker genes, highlighting enrichment for defense response activation, cytokine production, and leukocyte migration. E Representative images of CD68 and S100A12 immunofluorescence staining in knee synovial tissues from RA and OA patients. Scale bar: 100 μm

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Expressing, Marker, Activation Assay, Migration, Immunofluorescence, Staining

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: IRF7 is a specific transcriptional regulator of CD48 high S100A12 + macrophages. A Venn diagram showing overlapping transcription factors (TFs) identified by triplicate SCENIC analyses, with the CD48 high S100A12 + subcluster enriched for NFIL3, TGIF1, FOSL2, IRF7, and STAT1. B Heatmap of regulon activity scores (RASs) for TFs across macrophage subclusters. C Ranking of TFs in CD48 high S100A12 + macrophages by the regulon specificity score (RSS, calculated via Jensen‒Shannon divergence). D UMAP dimensionality reduction of TF activity profiles across subclusters. E – F UMAP plots highlighting spatial overlap between the CD48 high S100A12 + subcluster. ( E ) and cells with elevated IRF7 regulon activity ( F ). G Representative images of immunofluorescence staining for CD68 and IRF7 in knee synovial tissues from RA and OA patients. Scale bar: 100 μm

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Activity Assay, Immunofluorescence, Staining

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: IRF7 directly regulates downstream inflammatory genes in M1 macrophages. A ChIP-seq peak heatmaps showing increased IRF7 binding to promoter/enhancer regions in LPS-stimulated M1 macrophages. B Venn diagram of 108 overlapping genes from the IRF7 ChIP-seq data and the SCENIC-predicted target genes. C Reactome pathway enrichment of IRF7-regulated genes, highlighting the involvement of NF-κB, TNF, and Toll-like receptor signalling (key genes: IL-1β, FOS, NF-κB1, PTGS2, and CXCL10). D Bulk RNA-seq heatmap showing the upregulation of IRF7 and target genes in M1-polarized macrophages ( GSE130011 , GSE154346 ). E UMAP plots of NFKB1, PTGS2, IL1B, and CXCL10 expression in the CD48 high S100A12 + subcluster. F RT‒qPCR analysis of IRF7 and M1 marker genes in siRNA-treated macrophages (performed in triplicate, with 3 distinct patient sources used for each repetition). G – H Western blot validation of IRF7 and downstream protein expression following IRF7 knockdown in M1-polarized macrophages (performed in triplicate, with 3 distinct patient sources used for each repetition). Statistical significance: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way ANOVA with the Bonferroni post hoc correction)

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: ChIP-sequencing, Binding Assay, RNA Sequencing, Expressing, Marker, Western Blot, Biomarker Discovery, Knockdown

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: Local IRF7 knockdown alters the immune cell composition in CIA mice. A Schematic of intra-articular IRF7 siRNA treatment in collagen-induced arthritis (CIA) model mice. B – C Flow cytometry analysis of the CD86 and CD206 mean fluorescence intensities (MFIs) in F4/80 + macrophages from ankle joints ( n = 6). CD86: NC: 1241 ± 265.4, si-IRF7: 2469 ± 390.3, positive: 3489 ± 570.9, si-mock: 3689 ± 370.1. CD206: NC: 2225 ± 225.9, si-IRF7: 3395 ± 369.4, positive: 978.2 ± 147.9, si-mock: 2022 ± 170.6. D – E Frequencies of Foxp3 + Tregs among CD3 + CD4 + T cells ( n = 6). NC: 1.66% ± 0.15%, si-IRF7: 3.27% ± 0.28%, positive: 0.47% ± 0.17%, si-mock: 1.10% ± 0.22%. F Immunofluorescence staining for S100A12 + inflammatory macrophages in the ankle synovium of different groups. Scale bar: 100 μm. The data are presented as the means ± SDs. Statistical significance: *** P < 0.001, **** P < 0.0001 (one-way ANOVA with Bonferroni post hoc correction)

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Knockdown, Flow Cytometry, Fluorescence, Immunofluorescence, Staining

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: Local IRF7 inhibition attenuates joint inflammation and bone erosion in CIA mice. A Representative ankle joint images on day 42 postimmunization. B H&E staining and histological staining. C - E IHC staining for CD68, S100A12, and IRF7 in the ankle synovium. Scale bar: 100 μm. F Quantification of paw thickness at the ankle joint ( n = 6 per group). 42 Days after the first immunization: NC: 8.33 ± 0.02, si-IRF7: 10.39 ± 0.54, positive: 12.08 ± 0.80, si-mock: 12.65 ± 0.57, Statistical significance: **** P < 0.0001 (one-way ANOVA with Bonferroni post hoc correction). G H&E staining and histological scoring of synovial hyperplasia and inflammation ( n = 6). NC: 0.00 (0.00–0.00), si-IRF7: 1.50 (1.00–2.25), positive: 2.50 (1.75–3.00), and si-mock: 3.00 (2.75–3.00). Data are shown as medians with 25% − 75% percentiles. Statistical significance: * P < 0.05 (Kruskal‒Wallis test, followed by post hoc Dunn’s test with Bonferroni correction for multiple comparisons). H - J Semiquantitative analysis analysis of CD68, S100A12, and IRF7 expression via IHC staining via ImageJ ( n = 6). Statistical significance: **** P < 0.0001 (one-way ANOVA with Bonferroni post hoc correction)

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Inhibition, Staining, Immunohistochemistry, Expressing

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: 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: 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: The Journal of international medical research

Article Title: Identification of osteoarthritis biomarkers by proteomic analysis of synovial fluid.

doi: 10.1177/030006051204000622

Figure Lengend Snippet: FIGURE 2: Western blotting analysis for S100 calcium binding protein A12 (S100A12) in synovial fluid from patients with osteoarthritis (A,B) or rheumatoid arthritis (C,D)

Article Snippet: Molecular weight and electric charge data for potential biomarker peaks were used to identify proteins via the SWISS-PROT database (http://www.uniprot.org/).19 Western blotting of pooled synovial fluid samples was performed as described previously,20,21 using mouse-antihuman S100A12 primary antibody (Novus Biologicals, Littleton, CO, USA) and rabbit antimouse secondary antibody.

Techniques: Western Blot, Binding Assay