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: 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

Journal: Clinical & Translational Immunology

Article Title: Blockade of IL‐33 signalling attenuates osteoarthritis

doi: 10.1002/cti2.1187

Figure Lengend Snippet: Increased levels of IL‐33 and ST2 in human and murine OA. IL‐33 concentration (pg mL −1 ) in (a) SF and (b) serum of non‐OA ( n = 12) and OA patients ( n = 12). (c, d) mRNA and ( e) protein expression of IL‐33 and ST2 in isolated human chondrocytes from non‐OA ( n = 12) and OA patients ( n = 12). (f, g) OARSI scoring of cartilage tissue of sham‐operated ( n = 20) or DMM‐operated ( n = 20) WT mice (12 weeks post‐surgery end timepoint). (h) IL‐33 concentration (pg mL −1 ) in SF of sham‐operated ( n = 12) or DMM‐operated ( n = 12) WT mice (12 weeks post‐surgery end timepoint). (i, j) mRNA expression of IL‐33 and ST2 in whole knee joints from sham‐operated ( n = 20) or DMM‐operated ( n = 20) WT mice (12 weeks post‐surgery end timepoint). All RT‐qPCR gene expressions were normalised to the endogenous level of 18 s in respective groups. Data are expressed as mean ± S.E.M. with unpaired 2‐tailed Student’s t ‐tests. n indicates the number of human specimens or mice per group. NS = non‐significant. P < 0.0001 is represented as ****.

Article Snippet: Samples were probed overnight with primary antibody: IL‐33 (~ 30 kDa; 1:100; Novus Biologicals, Littleton; NBP1‐75516), ST2 (~ 50 and ~ 60 kDa; 1:100; Novus Biologicals; NBP2‐53096), MMP‐13 (~ 60 kDa; 1:100; Abcam, Cambridge; ab39012), ADAMTS‐5 (~ 73 kDa; 1:100; Abcam; ab41037), COL2A1 (~ 141 kDa 1:1000; Sigma‐Aldrich; SAB4500366) and β‐actin (loading control) (~ 42 kDa; 1:20 000; Sigma‐Aldrich; A2228).

Techniques: Concentration Assay, Expressing, Isolation, Quantitative RT-PCR

Journal: Clinical & Translational Immunology

Article Title: Blockade of IL‐33 signalling attenuates osteoarthritis

doi: 10.1002/cti2.1187

Figure Lengend Snippet: Neutralising ST2 attenuates OA. (a, b) OARSI scoring of cartilage tissue, (c) synovitis scoring and (d) osteophyte maturity scoring of sham‐operated ( n = 20) or DMM‐operated ( n = 20) WT mice (12 weeks post‐surgery end timepoint) treated intraperitoneally with either IgG1 (vehicle control; 50 µg per mouse; daily for 12 weeks post‐surgery) or αST2 (50 µg per mouse; daily for 12 weeks post‐surgery). (e) von Frey pain assessment of sham‐ ( n = 20) or DMM‐ ( n = 20) operated WT mice (12 weeks post‐surgery end timepoint) treated intraperitoneally with either IgG1 (vehicle control; 50 µg per mouse; daily for 12 weeks post‐surgery) or αST2 (50 µg per mouse; daily for 12 weeks post‐surgery). (f) mRNA expression of cartilage‐degrading proteases (MMP‐13, ADAMTS‐5 and MMP‐3) and chondrogenic markers (COL2A1, SOX‐9 and Aggrecan) in whole knee joints of DMM‐operated ( n = 20) treated intraperitoneally with either IgG1 (vehicle control; 50 µg per mouse; daily for 12 weeks post‐surgery) or αST2 (50 µg per mouse; daily for 12 weeks post‐surgery). (g) protein and (h) mRNA expression of cartilage degrading proteases (MMP‐13, ADAMTS‐5 and MMP‐3) and chondrogenic markers (COL2A1, SOX‐9 and Aggrecan) in isolated human chondrocytes from Non‐OA ( n = 20) and OA patients ( n = 20) treated with rIL‐33 (30 ng mL −1 , 24 h) and IgG1 (vehicle control; 3 μg mL −1 , 24 h) or αST2 (3 μg mL −1 , 24 h). (i) MMP‐13 (j) MMP‐3 protein level in cell media supernatants obtained from isolated human chondrocytes from Non‐OA ( n = 12) and OA patients ( n = 12) treated with rIL‐33 (30 ng mL −1 ; 24 h) and IgG1 (vehicle control; 3 μg mL −1 , 24 h) or αST2 (3 μg mL −1 , 24 h). All RT‐qPCR gene expressions were normalised to the endogenous level of 18 s in respective groups. Data are expressed as mean ± S.E.M. with two‐way analysis of variance followed by the Tukey‐Kramer test (b, c, d) or repeated measures 2‐way ANOVA with Bonferroni’s post hoc tests was used to compare groups at each time point ( e ; DMM IgG1 control mice vs DMM αST2 mice) or with unpaired 2‐tailed Student’s t ‐tests (i, j) . n indicates the number of human specimens or mice per group. NS = non‐significant. P < 0.001 or P < 0.0001 are represented as *** or ****, respectively.

Article Snippet: Samples were probed overnight with primary antibody: IL‐33 (~ 30 kDa; 1:100; Novus Biologicals, Littleton; NBP1‐75516), ST2 (~ 50 and ~ 60 kDa; 1:100; Novus Biologicals; NBP2‐53096), MMP‐13 (~ 60 kDa; 1:100; Abcam, Cambridge; ab39012), ADAMTS‐5 (~ 73 kDa; 1:100; Abcam; ab41037), COL2A1 (~ 141 kDa 1:1000; Sigma‐Aldrich; SAB4500366) and β‐actin (loading control) (~ 42 kDa; 1:20 000; Sigma‐Aldrich; A2228).

Techniques: Control, Expressing, Isolation, Quantitative RT-PCR

Journal: Clinical & Translational Immunology

Article Title: Blockade of IL‐33 signalling attenuates osteoarthritis

doi: 10.1002/cti2.1187

Figure Lengend Snippet: Blockade of IL‐33 signalling attenuates OA. IL‐33 is produced locally in OA joints by chondrocytes. Neutralising IL‐33 and ST2 antibodies attenuates the severity of OA and pain in vivo and decreases the production of cartilage degrading proteases.

Article Snippet: Samples were probed overnight with primary antibody: IL‐33 (~ 30 kDa; 1:100; Novus Biologicals, Littleton; NBP1‐75516), ST2 (~ 50 and ~ 60 kDa; 1:100; Novus Biologicals; NBP2‐53096), MMP‐13 (~ 60 kDa; 1:100; Abcam, Cambridge; ab39012), ADAMTS‐5 (~ 73 kDa; 1:100; Abcam; ab41037), COL2A1 (~ 141 kDa 1:1000; Sigma‐Aldrich; SAB4500366) and β‐actin (loading control) (~ 42 kDa; 1:20 000; Sigma‐Aldrich; A2228).

Techniques: Produced, In Vivo