Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: A Flow cytometry results of CD73-positive ADSCs. B Flow cytometry results of CD73-negative ADSCs. C In vitro, immunofluorescence staining of ADSCs in each group. Red indicates CD73, green indicates VEGF, blue indicates DAPI, scale bar = 20 µm. D Quantitative analysis of immunofluorescence staining results among different groups ( n = 3). E Western blot results of CD73 and VEGF proteins after transfection or APCP treatment in each group. F Quantitative analysis of Western blot results. G CCK-8 assay results among different groups of ADSCs in conditioned medium ( n = 3). H Colony formation assay results among RBSMC in conditioned medium ( n = 3). I Cell migration assay results and quantitative analysis among different groups of ADSCs in conditioned medium ( n = 3). J Wound healing assay results among RBSMC in conditioned medium ( n = 3). Data presented as ±SD. * and # indicates P < 0.05, ** and ## indicates P < 0.01, and *** and ### indicates P < 0.001. Full-length blots/gels are presented in Supplementary Fig. .

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Flow Cytometry, In Vitro, Immunofluorescence, Staining, Western Blot, Transfection, CCK-8 Assay, Colony Assay, Cell Migration Assay, Wound Healing Assay

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: A Western blot results of CD73 and VEGF in the bladder tissues of NB + CD73⁺ / ⁺ group on days 0, 7, 14, 21, and 28 after treatment. B Quantitative analysis of the Western blot results for CD73 and VEGF ( n = 3). C The results of the mean pressure of the voiding contractions and mean intermicturition interval in each group of rats ( n = 5). D Representative images of cystometrography results for each group of rats ( n = 5). E Masson staining results of bladder tissues in each group of rats. scale bar = 50 µm. F Immunofluorescence staining results for CD73 and VEGF in the bladder tissues of each group. Red indicates CD73, green indicates VEGF, and blue indicates DAPI. scale bar = 20 µm. G Quantitative analysis of smooth muscle content in the bladder (yellow square) of each group ( n = 3). H Quantitative analysis of immunofluorescence staining for CD73 and VEGF ( n = 3). Data presented as ±SD. * and # indicates P < 0.05, ** and ## indicates P < 0.01, and *** and ### indicates P < 0.001. Full-length blots/gels are presented in Supplementary Fig. .

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Western Blot, Staining, Immunofluorescence

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: A Immunofluorescence staining results of ADSCs in the bladder tissues of each group of rats. Red represents ADSCs, green represents βIII-tubulin, and blue represents DAPI. Scale bar = 20 µm. B Immunohistochemical results of CXCR4 in the bladder tissues of each group. The green arrows indicate regions of high CXCR4 expression. Scale bar = 50 µm. C Immunofluorescence staining results of CD73 and SDF-1 in the bladder tissues of each group. Red represents SDF-1, green represents CD73, and blue represents DAPI. Scale bar = 20 µm. D Quantitative results of ADSCs in the bladder tissues of each group ( n = 5). E Quantitative results of SDF-1 expression in the bladder tissues of each group ( n = 3). F In vitro, western blot results of SDF-1 in each group under conditioned medium. G Quantitative results of SDF-1 expression of each group ( n = 3). H Cell migration assay results of CD73⁺ ADSCs after SDF-1 gene knockdown. I Quantitative results of cell migration assays for ADSCs in each group ( n = 3). Data presented as ±SD. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001. Full-length blots/gels are presented in Supplementary Fig. . Full-section IHC are presented in Supplementary Fig. .

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Immunofluorescence, Staining, Immunohistochemical staining, Expressing, In Vitro, Western Blot, Cell Migration Assay, Knockdown, Migration

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: CD73 activation upregulates VEGF expression, further stimulating the PI3K/AKT/mTOR pathway to enhance cell proliferation. Simultaneously, it inhibits NFκB phosphorylation, suppressing the NFκB/NLRP3/caspase-1 axis, thereby preventing apoptosis and reducing IL-1β and IL-6 levels. Moreover, activated CD73 increases SDF-1 expression, which interacts with its receptor CXCR4 to direct cell migration to damaged bladder tissue.

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Activation Assay, Expressing, Phospho-proteomics, Migration

Journal: The Journal of biological chemistry

Article Title: Fibroblast growth factor-8 expression is regulated by intronic engrailed and Pbx1-binding sites.

doi: 10.1074/jbc.274.9.6020

Figure Lengend Snippet: FIG. 6. En1, En2, and Pbx1 transcription factors present in day 9 embryoid bodies. Western blots of nuclear extracts, probed with antisera against Engrailed proteins (left panel) or Pbx1 (right panel). The 55-kDa En1 and 41-kDa En2 proteins are indicated in the left blot, while the 53-kDa Pbx1 protein is indicated in the right blot. The protein molecular weight sizes are indicated in kilodaltons between the blots.

Article Snippet: Western blots on nuclear extracts were performed as described previously (35), using a primary antibody to En1 and En2 (36), or a primary antibody to Pbx1 (Santa Cruz Biotechnology, Inc.).

Techniques: Western Blot, Molecular Weight

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: 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

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