Journal: The journal of applied laboratory medicine

Article Title: Plasma Protein Profiling by Proximity Extension Assay Technology Reveals Novel Biomarkers of Traumatic Brain Injury-A Pilot Study.

doi: 10.1093/jalm/jfab004

Figure Lengend Snippet: Fig. 1. Normalized protein expression (NPX) of plasma biomarkers in surviving [sTBI (S); n 5 6] and non- surviving sTBI patients [sTBI (NS); n 5 4] compared with matched healthy controls (CTR, n 5 10). Biomarkers shown are (A) ST2, (B) IL6, (C) TNNI3, (D) TNFR-2, (E) CHI3L1, and (F) EN-RAGE. Reference groups shown are ovarian cancer patients (OvCa, n 5 32) and dementia patients (mild cognitive impair- ment, MCI, n 5 18; Alzheimer disease, AD, n 5 12). Dotted lines represent group median, bars show interquartile range. Mann Whitney U test; *P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant. For more discussion see text.

Article Snippet: All sTBI and matched control samples were analyzed in duplicate using the commercially available ELISA kits, DRT200 TNFR2 Quantikine kit and the DST200 ST2 Quantikine kit (R&D Systems), according to manufacturer’s instructions.

Techniques: Expressing, Clinical Proteomics, MANN-WHITNEY

Journal: The journal of applied laboratory medicine

Article Title: Plasma Protein Profiling by Proximity Extension Assay Technology Reveals Novel Biomarkers of Traumatic Brain Injury-A Pilot Study.

doi: 10.1093/jalm/jfab004

Figure Lengend Snippet: Fig. 3. Plasma concentration of ST2 and TNFR-2 in healthy controls (n 5 10) and sTBI patients (n 5 10), as measured by commercially available ELISAs. Mann Whitney U test; ****P < 0.0001. For more details see text.

Article Snippet: All sTBI and matched control samples were analyzed in duplicate using the commercially available ELISA kits, DRT200 TNFR2 Quantikine kit and the DST200 ST2 Quantikine kit (R&D Systems), according to manufacturer’s instructions.

Techniques: Clinical Proteomics, Concentration Assay, MANN-WHITNEY

Journal: The journal of applied laboratory medicine

Article Title: Plasma Protein Profiling by Proximity Extension Assay Technology Reveals Novel Biomarkers of Traumatic Brain Injury-A Pilot Study.

doi: 10.1093/jalm/jfab004

Figure Lengend Snippet: Fig. 4. Comparison between commercially available proximity extension assays (Olink Proteomics) and ELISAs. Plasma ST2 was measured in (A) healthy controls (n 5 10); Spearman’s correlation (rs) 5 0.8909, P 5 0.0011, and (B) sTBI patients (n 5 10); rs 5 0.9758, P < 0.0001. Plasma TNFR-2 was measured in (A) healthy controls (n 5 10); rs 5 0.9030, P 5 0.0008, and (B) sTBI patients (n 5 10); rs 5 0.8667, P 5 0.0022. Black line shows line of best fit.

Article Snippet: All sTBI and matched control samples were analyzed in duplicate using the commercially available ELISA kits, DRT200 TNFR2 Quantikine kit and the DST200 ST2 Quantikine kit (R&D Systems), according to manufacturer’s instructions.

Techniques: Comparison, Clinical Proteomics

Journal: Cell reports

Article Title: Organoids capture tissue-specific innate lymphoid cell development in mice and humans.

doi: 10.1016/j.celrep.2022.111281

Figure Lengend Snippet: Figure 3. SIOs provide an essentially GF model of gut-specific ILCs (A) Representative flow plots of NKp46 expression in ILCP + SIO co-culture-derived ILCs or SI-LP-derived CD127+ ILCs, with the frequency of NKp46+ ILC3s (co- culture: Live, EpCAM, Lin, CD45+, RORgt+; primary tissue: Live, CD45+, Lin, CD127+, Klrg1, NK1.1+/, RORgt+) additionally quantified for ILCPs cultured without SIOs or with GF SIOs in (B) (N = 2–5 animals, pooled from two experiments). (C) Relative frequency of mature ILC subsets excluding immature or other cells, depicting group 1 (magenta; Live, EpCAM, CD45+, Lin, RORgt-, ST2, Klrg1, NK1.1+, NKp46+), group 2 (green; Live, EpCAM, CD45+, Lin, RORgt, NK1.1, ST2+, Klrg1+, Sca-1+), NKp46+ group 3 (lavender; Live, EpCAM, CD45+, Lin, ST2, Klrg1, RORgt+, NKp46+), and NKp46 group 3 ILCs (blue; Live, EpCAM, CD45+, Lin, ST2, Klrg1, RORgt+, NKp46) in live, unstimulated co-cultures derived from SPF-SIOs or GF-SIOs compared with primary SPF ileum (no Peyer’s patches). (D) Diagram of transwell culture strategy. (E) Relative frequency of group 1, 2, and 3 ILCs derived from PD-1+ ILCP + SIO +/ transwell insert (TW) separation (N = 3, two experiments).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Human CRTh2 – PE Miltenyi Biotec Cat# 130-113-600 Human CRTh2– BV711 BioLegend Cat# 350124 Human CRTh2 – BV421 BioLegend Cat# 350112 Human CD161 APC, A700 BioLegend Cat# 302012 Human ST2 – APC R&D Sys Cat# FAB5231A Human ST2 – PE R&D Sys Cat# FAB5231P Human NKp44 – PE Cy7 BioLegend Cat# 325116 Human NKp44 – PerCP Cy5.5 BioLegend Cat# 325114 Human/Mouse Tbet – PE-Cy7 BioLegend Cat# 644824 Human RORgt – APC Thermofisher (eBioscience) Cat# 17-6988-82 Human RORgt – PE BDBiosciences Cat# 563081 Human GATA3 – APC Cy7 Santa Cruz Cat# sc-268 Human IL22 – PerCP Cy5.5 BioLegend Cat# 366709 Human IL17A – PE-Dazzle BioLegend Cat# 512335 Human IL17A – e450 BD Horizon/ bioscience Cat# 560610 Human IL17A – BV786 BD Horizon/ bioscience Cat# 563745 Human IFNg –APCe780 Invitrogen Cat# 47-7319-41 Human IL-5 – APC BioLegend Cat# 504305 Human IL-13 – FITC eBioscience Cat# 11-7139-41 Human IL-13 – bv711 BD Biosciences Cat# 564288 Human CD25 – PerCP-Cy 5.5 BD Biosciences Cat# 560503 Human Klrg1 – APC Thermofisher (eBioscience) Cat# 25-5893-80 Human CCR6 – APC BioLegend Cat# 353416 Human CCR6 – BV605 BioLegend Cat# 353419 Human NKp46 APC BioLegend Cat# 331918 FcR CD16/32 blocking, mouse BioCell Cat# BE0307 FcR blocking, human Miltenyi Biotec Cat# 130-059-901 Anti-rhIL33 (neutralising, ICC, goat polyclonal) R&D Cat# AF3625 Anti-rmIL33 (neutralising, ICC, goat polyclonal) R&D Cat# AF3626 E-Cadherin – anti-human (rat) Thermofisher (eBioscience) Cat# 51-3249-82 CDX2 – anti-human (rabbit) abcam Cat# Ab76541 EpCAM – anti-mouse (rabbit) abcam Cat# Ab71916 CD45 anti-human (mouse) BioLegend Cat# 304001 ZO-1 anti-mouse (rabbit) Abcam Cat# Ab96587 Dclk1 anti-mouse (rabbit) Abcam Cat# Ab31704 CD44 anti-mouse/human (rat) Thermofisher (eBioscience) Cat# 14-0551-82 Critical commercial assays CellTrace FarRed ThermoFischer Cat# C34564 Foxp3 / Transcription Factor Staining Buffer Set Invitrogen eBioscience Cat# 00-5523-00 Live Dead fixable blue/UV ThermoFischer Cat# L34961 UltraComp eBeads Invitrogen Cat# 01-2222-42 RNeasy Qiagen Cat# 74106 RevertAid First Strand cDNA Synthesis Kit ThermoFisher Cat# K1622 Fast SYBR green master mix Applied Biosystems Cat# 4385612 (Continued on next page) Cell Reports 40, 111281, August 30, 2022 e2

Techniques: Expressing, Co-Culture Assay, Derivative Assay, Cell Culture

Journal: Cell reports

Article Title: Organoids capture tissue-specific innate lymphoid cell development in mice and humans.

doi: 10.1016/j.celrep.2022.111281

Figure Lengend Snippet: Figure 7. Gut-matured ILC2 upregulates ST2 on transfer to HLO culture (A) Representative image of SD-HIOs and SD-HLOs showing E-cadherin+ (E-CAD) epithelium, CD45+ ILCs, and nuclei (Hoechst) after 14-day co-culture (scale bars: 50 mm). (B) Count of EpCAM, CD45+, LIN ILCs after 14-day co-culture with SD-HIOs or SD-HLOs, with corresponding count of EpCAM, CD45 mesenchyme.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Human CRTh2 – PE Miltenyi Biotec Cat# 130-113-600 Human CRTh2– BV711 BioLegend Cat# 350124 Human CRTh2 – BV421 BioLegend Cat# 350112 Human CD161 APC, A700 BioLegend Cat# 302012 Human ST2 – APC R&D Sys Cat# FAB5231A Human ST2 – PE R&D Sys Cat# FAB5231P Human NKp44 – PE Cy7 BioLegend Cat# 325116 Human NKp44 – PerCP Cy5.5 BioLegend Cat# 325114 Human/Mouse Tbet – PE-Cy7 BioLegend Cat# 644824 Human RORgt – APC Thermofisher (eBioscience) Cat# 17-6988-82 Human RORgt – PE BDBiosciences Cat# 563081 Human GATA3 – APC Cy7 Santa Cruz Cat# sc-268 Human IL22 – PerCP Cy5.5 BioLegend Cat# 366709 Human IL17A – PE-Dazzle BioLegend Cat# 512335 Human IL17A – e450 BD Horizon/ bioscience Cat# 560610 Human IL17A – BV786 BD Horizon/ bioscience Cat# 563745 Human IFNg –APCe780 Invitrogen Cat# 47-7319-41 Human IL-5 – APC BioLegend Cat# 504305 Human IL-13 – FITC eBioscience Cat# 11-7139-41 Human IL-13 – bv711 BD Biosciences Cat# 564288 Human CD25 – PerCP-Cy 5.5 BD Biosciences Cat# 560503 Human Klrg1 – APC Thermofisher (eBioscience) Cat# 25-5893-80 Human CCR6 – APC BioLegend Cat# 353416 Human CCR6 – BV605 BioLegend Cat# 353419 Human NKp46 APC BioLegend Cat# 331918 FcR CD16/32 blocking, mouse BioCell Cat# BE0307 FcR blocking, human Miltenyi Biotec Cat# 130-059-901 Anti-rhIL33 (neutralising, ICC, goat polyclonal) R&D Cat# AF3625 Anti-rmIL33 (neutralising, ICC, goat polyclonal) R&D Cat# AF3626 E-Cadherin – anti-human (rat) Thermofisher (eBioscience) Cat# 51-3249-82 CDX2 – anti-human (rabbit) abcam Cat# Ab76541 EpCAM – anti-mouse (rabbit) abcam Cat# Ab71916 CD45 anti-human (mouse) BioLegend Cat# 304001 ZO-1 anti-mouse (rabbit) Abcam Cat# Ab96587 Dclk1 anti-mouse (rabbit) Abcam Cat# Ab31704 CD44 anti-mouse/human (rat) Thermofisher (eBioscience) Cat# 14-0551-82 Critical commercial assays CellTrace FarRed ThermoFischer Cat# C34564 Foxp3 / Transcription Factor Staining Buffer Set Invitrogen eBioscience Cat# 00-5523-00 Live Dead fixable blue/UV ThermoFischer Cat# L34961 UltraComp eBeads Invitrogen Cat# 01-2222-42 RNeasy Qiagen Cat# 74106 RevertAid First Strand cDNA Synthesis Kit ThermoFisher Cat# K1622 Fast SYBR green master mix Applied Biosystems Cat# 4385612 (Continued on next page) Cell Reports 40, 111281, August 30, 2022 e2

Techniques: Co-Culture Assay

Journal: International journal of molecular medicine

Article Title: Nrf2 activation diminishes during adipocyte differentiation of ST2 cells.

doi: 10.3892/ijmm.2011.761

Figure Lengend Snippet: Figure 1. The intracellular glutathione redox potential becomes more oxidized during adipocyte differentiation of ST2 cells. ST2 cells stably expressing Grx1- roGFP2 were induced to differentiate into adipocytes. Every 24 h the cells were excited with 390 and 480 nm lasers and the ratio of emissions in the green channel (535 nm) was calculated. The graph represents the mean ± SD. Three independent experiments were performed, and in each experiment 24 technical replicates were used for each timepoint. *p<0.0001 compared to day 0; †p<0.001; ††p<0.0001.

Article Snippet: ST2 cells are a cloned stromal-cell line from mouse bone marrow; obtained from DSMZ (German Collection of Microorganisms and Cell Cultures).

Techniques: Stable Transfection, Expressing

Journal: International journal of molecular medicine

Article Title: Nrf2 activation diminishes during adipocyte differentiation of ST2 cells.

doi: 10.3892/ijmm.2011.761

Figure Lengend Snippet: Figure 2. Nuclear Nrf2 protein abundance decreases in ST2 cells during adi- pocyte differentiation. ST2 cells were induced to differentiate into adipocytes. Nuclear protein extracts were prepared immediately before the initiation of dif- ferentiation (day 0) and every 24 h thereafter until day 5. Nrf2 expression was assayed by Western immunoblotting. The nuclear abundance of the transcrip- tion factor SP-1 was employed as a loading control. The image is representative of 3 independent experiments. The graph represents the mean ± SD. *p<0.0001 compared to day 0. †p<0.05; ††p<0.0001.

Article Snippet: ST2 cells are a cloned stromal-cell line from mouse bone marrow; obtained from DSMZ (German Collection of Microorganisms and Cell Cultures).

Techniques: Quantitative Proteomics, Expressing, Western Blot, Control

Journal: International journal of molecular medicine

Article Title: Nrf2 activation diminishes during adipocyte differentiation of ST2 cells.

doi: 10.3892/ijmm.2011.761

Figure Lengend Snippet: Figure 3. Nrf2 binding activity decreases during adipocyte differentiation of ST2 cells. ST2 cells were induced to differentiate into adipocytes. Nuclear protein extracts were prepared before the initiation of differentiation (day 0) and every 24 h thereafter until day 5, and were used for EMSAs with an ARE- containing oligonucleotide as a radiolabelled probe. The image is representative of 3 independent experiments. Binding competition experiments performed with ST2 nuclear extracts at day 0 (0d) of differentiation (maximal Nrf2 DNA binding activity) in the presence of a 10-, 50-, or 100-fold molar excess of unlabeled ARE-wt or ARE-mut oligonucleotides. The image is representative of 3 independent experiments.

Article Snippet: ST2 cells are a cloned stromal-cell line from mouse bone marrow; obtained from DSMZ (German Collection of Microorganisms and Cell Cultures).

Techniques: Binding Assay, Activity Assay

Journal: International journal of molecular medicine

Article Title: Nrf2 activation diminishes during adipocyte differentiation of ST2 cells.

doi: 10.3892/ijmm.2011.761

Figure Lengend Snippet: Figure 4. ARE-mediated transcription is reduced during differentiation of ARE-ST2 cells into adipocytes. ARE-ST2 cells were induced to differentiate into adipocytes and luciferase measurements were performed every 24 h until day 5 of differentiation. Luciferase activity was normalized to total protein concentration. The graph represents the means ± SD. Three independent exper- iments were performed, and in each experiment 8 technical replicates were used for each timepoint. *p<0.0001 compared to day 0; †p<0.05; ††p<0.0001.

Article Snippet: ST2 cells are a cloned stromal-cell line from mouse bone marrow; obtained from DSMZ (German Collection of Microorganisms and Cell Cultures).

Techniques: Luciferase, Activity Assay, Protein Concentration

Journal: BMC pharmacology & toxicology

Article Title: Blocking group 2 innate lymphoid cell activation and macrophage M2 polarization: potential therapeutic mechanisms in ovalbumin-induced allergic asthma by calycosin.

doi: 10.1186/s40360-024-00751-9

Figure Lengend Snippet: Fig. 2 Calycosin inhibited group 2 innate lymphoid cell (ILC2) activation in OVA-induced asthmatic mice. (A) The levels of interleukin (IL)-4, (B) IL-5 and (C) IL-13 in the BALF and lung tissues were measured using ELISA and RT-qPCR. (D) ELISA was used to determin the contents of IL-33 in the BALF. (E) Suppression tumorigenicity 2 (ST2) levels in the lung tissues was examined by western blot. The blots were cut prior to hybridization with antibodies during blotting. (F) Immunohistochemistry (IHC) staining was used to detect the expression ST2 and IL-25R. (G) The co-localization of ST2 and CD45 was detected by immunofluorescence (IF). Scale bar = 50 μm. n = 6 for each group. * P< 0.05, **P < 0.01, ***P < 0.001, compared with the sham group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the OVA group. “Ca” represents calycosin

Article Snippet: The sections were incubated with ST2 antibody (1:50; 11920- 1-AP, ProteinTech, Wuhan, China) at 4°C overnight and subjected to HRP-labeled goat anti-rabbit IgG (1:500; # 31460, ThermoFisher Scientific, USA) at 37°C for 1 h. Additionally, IL-25R expression was detected in the same way.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Hybridization, Immunohistochemistry, Expressing, Immunofluorescence

Journal: BMC pharmacology & toxicology

Article Title: Blocking group 2 innate lymphoid cell activation and macrophage M2 polarization: potential therapeutic mechanisms in ovalbumin-induced allergic asthma by calycosin.

doi: 10.1186/s40360-024-00751-9

Figure Lengend Snippet: Fig. 4 ILC2 activation induced by IL-33 was restrained by calycosin in ILC2s. (A) The ILC2 cell viability was examined by CCK-8. (B) Western blot showed the expression of ST2 in ILC2s. The blots were cut prior to hybridization with antibodies during blotting. (C) The secretion of IL-4, (D) IL-5 and (E) IL-13 in ILC2s were detected by ELISA assay. n = 3 for each group. **P < 0.01,***P < 0.001, compared with the control group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the IL-33 group. “Ca” represents calycosin

Article Snippet: The sections were incubated with ST2 antibody (1:50; 11920- 1-AP, ProteinTech, Wuhan, China) at 4°C overnight and subjected to HRP-labeled goat anti-rabbit IgG (1:500; # 31460, ThermoFisher Scientific, USA) at 37°C for 1 h. Additionally, IL-25R expression was detected in the same way.

Techniques: Activation Assay, CCK-8 Assay, Western Blot, Expressing, Hybridization, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Clinical Medicine

Article Title: Dynamics of the Novel Cardiac Biomarkers sST2, H-FABP, GDF-15 and suPAR in HFrEF Patients Undergoing Heart Failure Therapy, a Pilot Study

doi: 10.3390/jcm14165668

Figure Lengend Snippet: Serum concentrations before and after ARNI therapy of ( A ) sST2, ( B ) H-FABP, ( C ) GDF-15, and ( D ) suPAR. *: p < 0.05.

Article Snippet: Serum concentrations of sST2 (DY523B-05), suPAR (DY807), GDF-15 (DY957) and H-FABP (DY1678) were measured by commercially available enzyme-linked immunosorbent assays (ELISAs), purchased from R & D Systems (Abingon, UK).

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Dynamics of the Novel Cardiac Biomarkers sST2, H-FABP, GDF-15 and suPAR in HFrEF Patients Undergoing Heart Failure Therapy, a Pilot Study

doi: 10.3390/jcm14165668

Figure Lengend Snippet: Correlation of change in serum concentration of ( A ) sST2 and NT-proBNP, ( B ) GDF-15 and NT-proBNP, ( C ) sST2 and GDF-15, and ( D ) GDF-15 and ESC Score 2.

Article Snippet: Serum concentrations of sST2 (DY523B-05), suPAR (DY807), GDF-15 (DY957) and H-FABP (DY1678) were measured by commercially available enzyme-linked immunosorbent assays (ELISAs), purchased from R & D Systems (Abingon, UK).

Techniques: Concentration Assay