Journal: Arthritis Research & Therapy

Article Title: The role of interleukin-1 in the pathogenesis of human Intervertebral disc degeneration

doi: 10.1186/ar1732

Figure Lengend Snippet: Real-time PCR probes and details of primers

Article Snippet: Sections were incubated overnight at 4°C with mouse monoclonal primary antibodies against human IL-1Ra (1:200 dilution, R&D Systems, Abingdon, UK), IL-1RI (1:50 dilution, R&D Systems), and goat polyclonal primary antibodies against human IL-1α (1:300 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-1β (1:300 dilution, SantaCruz), and ICE (1:10 dilution, SantaCruz).

Techniques: Real-time Polymerase Chain Reaction

Journal: Arthritis Research & Therapy

Article Title: The role of interleukin-1 in the pathogenesis of human Intervertebral disc degeneration

doi: 10.1186/ar1732

Figure Lengend Snippet: Examples of imunohistochemical staining for the IL-1 family. IL-1β (row A), IL-1Ra (row B), and IL-1 receptor, type I (row C) in grade-1 non-degenerate discs (column 1) and grade-12 degenerate discs (column 2), IgG controls (row D) were all negative. Immunopositivity is revealed by brown staining. N.B In non-degenerate discs, no cell clusters were seen and little immunopositivity was observed in the single cells. In degenerate discs, a large number of cell clusters were observed, which were predominately immunopositive. Bars = 570 μm.

Article Snippet: Sections were incubated overnight at 4°C with mouse monoclonal primary antibodies against human IL-1Ra (1:200 dilution, R&D Systems, Abingdon, UK), IL-1RI (1:50 dilution, R&D Systems), and goat polyclonal primary antibodies against human IL-1α (1:300 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-1β (1:300 dilution, SantaCruz), and ICE (1:10 dilution, SantaCruz).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: The role of interleukin-1 in the pathogenesis of human Intervertebral disc degeneration

doi: 10.1186/ar1732

Figure Lengend Snippet: Immunopositive staining for the IL-1 family in human intervertebral discs. Numbers of cells with immunopositivity for IL-1α (a) , IL-1β (b) , IL-1 receptor antagonist (c) , IL-1 receptor, type I (d) , and IL-1β-converting enzyme (e) , according to place of origin in the disc and grade of intervertebral disc degeneration ( n = 30). Data are presented as means ± 2 standard errors (as a representative of 95%CI). * P < 0.1,; ** P < 0.05

Article Snippet: Sections were incubated overnight at 4°C with mouse monoclonal primary antibodies against human IL-1Ra (1:200 dilution, R&D Systems, Abingdon, UK), IL-1RI (1:50 dilution, R&D Systems), and goat polyclonal primary antibodies against human IL-1α (1:300 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-1β (1:300 dilution, SantaCruz), and ICE (1:10 dilution, SantaCruz).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: The role of interleukin-1 in the pathogenesis of human Intervertebral disc degeneration

doi: 10.1186/ar1732

Figure Lengend Snippet: Effects of IL-1β on gene expression in cells from non- degenerate or degenerate intervertebral discs

Article Snippet: Sections were incubated overnight at 4°C with mouse monoclonal primary antibodies against human IL-1Ra (1:200 dilution, R&D Systems, Abingdon, UK), IL-1RI (1:50 dilution, R&D Systems), and goat polyclonal primary antibodies against human IL-1α (1:300 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-1β (1:300 dilution, SantaCruz), and ICE (1:10 dilution, SantaCruz).

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: The role of interleukin-1 in the pathogenesis of human Intervertebral disc degeneration

doi: 10.1186/ar1732

Figure Lengend Snippet: Effect of IL-1 treatment on the IL-1 family gene expression in human intervertebral disc cells. Relative gene expression was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeping gene and untreated controls (hence control is graphed at 1 on the log scale) for IL-1α (a) , IL-1β (b) , IL-1 receptor antagonist (IL-1Ra) (c) , and IL-1 receptor, type I (IL-RI) (d) following IL-1β treatment of disc cells from two regions of non-degenerate (non-deg) ( n = 6) and degenerate ( n = 24) discs. ** P < 0.05.

Article Snippet: Sections were incubated overnight at 4°C with mouse monoclonal primary antibodies against human IL-1Ra (1:200 dilution, R&D Systems, Abingdon, UK), IL-1RI (1:50 dilution, R&D Systems), and goat polyclonal primary antibodies against human IL-1α (1:300 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-1β (1:300 dilution, SantaCruz), and ICE (1:10 dilution, SantaCruz).

Techniques: Expressing, Control

Journal: Nature Communications

Article Title: Inhibition of interleukin-1β reduces myelofibrosis and osteosclerosis in mice with JAK2 -V617F driven myeloproliferative neoplasm

doi: 10.1038/s41467-022-32927-4

Figure Lengend Snippet: a Upper panel: Serum IL-1β (pg/ml) in normal controls (NC; n = 20) and MPN patients ( n = 120); ET ( n = 42), PV ( n = 44), PMF ( n = 34). Correlation ( r ) between % JAK2 -V617F in granulocytes and log transformed serum IL-1β in MPN patients. Limit of detection is shown by dashed green line at y = 0.01 pg/ml. Lower panel: IL-1β mRNA expression relative to β-actin in granulocytes of NC ( n = 12) and MPN patients ( n = 46); ET ( n = 11), PV ( n = 18), PMF ( n = 17). Correlation between log transformed IL1B mRNA expression and % JAK2 -V617F. b Upper panel: Serum IL-1RA (pg/ml) in NC ( n = 20) and MPN patients ( n = 120); ET ( n = 42), PV ( n = 44), PMF ( n = 34). Correlation ( r ) between % JAK2 -V617F and log transformed serum IL-1RA. Lower panel: IL1RN (IL-1RA) mRNA expression relative to β-actin in NC and MPN patients. Correlation between log transformed IL1RN mRNA expression and % JAK2 -V617F. Two-tailed unpaired non-parametric Mann–Whitney t -test was performed in a and b . c Representative histogram showing the expression of interleukin 1 receptor type 1 (IL-1R1) in peripheral blood hematopoietic stem cells (HSCs) from isotype control, NC ( n = 5), ET ( n = 6), PV ( n = 5), and PMF ( n = 7). Bar graph showing the percentages of IL-1R1 + HSC), common myeloid progenitors (CMP), granulocyte macrophage progenitor (GMP), megakaryocyte erythroid progenitor (MEP) and megakaryocyte progenitor (MkP). Graph showing correlation ( r ) between % JAK2 -V617F and percentages of IL-1R1 + HSCs. d Representative histogram showing the expression of interleukin 1 receptor accessory protein (IL-1RAcP) in peripheral blood HSC from NC and MPN patients. Bar graph showing the percentages of IL1RAcP + HSC, CMP, GMP, MEP, and MkP in NC ( n = 5), ET ( n = 6), PV ( n = 5), and PMF ( n = 7). Correlation ( r ) between % JAK2 -V617F and percentages of IL-1RAcP+ HSPCs. Two-tailed unpaired t-test was performed for statistical comparisons in c and d . Spearman correlation ( r ) and two-tailed t -test was performed for correlation analysis in a – d . All data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. See also Supplementary Figs. – . Source data and exact p values are provided as a Source Data file.

Article Snippet: Frozen PBMCs from MPN patients and normal controls were thawed and stained after blocking Fcγ receptors (#564220, BD) with following human antibodies: lineage-FITC (1:20; 348701), CD34-Pacific Blue (1:100; 343512), CD38-APC (1:50; 356606), CD123-BV605 (1:100; 306026), and CD41-PE-Cy5 (1:50; 343512) from BioLegend, CD45RA-BV786, (1:50; 563870; BD biosciences) and IL-1R1-PE (1:20; FAB269P), IL-1RAcP-PE (1:20; FAB676P) or isotype goat IgG-PE antibody (1:20; IC108P) from R&D systems.

Techniques: Transformation Assay, Expressing, Two Tailed Test, MANN-WHITNEY, Control

Journal: Frontiers in Immunology

Article Title: Siponimod (BAF312) Activates Nrf2 While Hampering NFκB in Human Astrocytes, and Protects From Astrocyte-Induced Neurodegeneration

doi: 10.3389/fimmu.2020.00635

Figure Lengend Snippet: Characterization of iPSC-derived astrocytes. (A) Phase contrast images showing human iNPC (left panel) and human iAstrocytes (right panel). (B,C) Representative immunofluorescence stainings for GFAP, S100β, nestin, vimentin (B) and S1P1, S1P3, IL1R and IL17R (C) in human iAstrocytes. DAPI was used for nuclear staining. (D,E) Percentage of cells positive for astrocyte markers at two timepoints during differentiation. Reported quantifications were performed on three different human iAstrocytes preparations from the same iNPC cell line. Bars represent SEM. Same observations were recorded in human iAstrocytes from a second iNPC cell line. Scale bar = 30 μm.

Article Snippet: The following primary antibodies were used: rabbit anti-GFAP (Agilent), mouse anti-nestin (Merck Millipore), mouse anti-vimentin (Abcam), rabbit anti-S100β (Abcam), rabbit anti-EDG1 (Santa Cruz Biotechnology), rabbit anti-EDG3 (Santa Cruz Biotechnology), mouse anti-IL1R (R&D), rabbit anti-IL17R (Santa Cruz biotechnology), rabbit anti- NFκB p65 (Abcam), rabbit anti-GLAST (Abcam), guinea pig anti-GLT1 (Merck Millipore), rabbit anti-Nrf2 (Abcam), mouse anti Neuronal Class III β-Tubulin (Covance).

Techniques: Derivative Assay, Immunofluorescence, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: IL-1beta mediates the proangiogenic activity of osteopontin-activated human monocytes.

doi: 10.4049/jimmunol.177.7.4267

Figure Lengend Snippet: FIGURE 3. OPN-treated monocytes induce endothelial cell chemotaxis and angiogenesis in the CAM through IL-1. A, Murine aortic endothelial cells were assessed for their capacity to migrate in response to CM from control and OPN-treated monocytes in a Boyden chamber assay. After 4 h, cells that migrated through the filter were counted in triplicate (five fields per well) at 250 magnification. Control values obtained in fresh medium (52 8 cells/field) were subtracted from all the data. , Significantly different from control CM, p 0.01 by Student’s t test. B, Gelatin sponges were adsorbed with: vehicle (PBS); rFGF-2; CM from OPN-treated mono- cytes in the absence (CM) or in the presence of neutralizing anti-IL-1 (CM anti-IL-1), anti-IL-8 (CM anti-IL-8), or anti-TNF- (CM anti-TNF-) mAb; and rIL-1. Sponges were then implanted onto the CAM. At day 12, blood vessels entering the sponges were counted. Data are the mean SD of 20 embryos. , Significantly different from CM, p 0.05 by Student’s t test. C–E, Macroscopic images of CAM treated with CM from OPN-treated monocytes in the absence (C) or in the presence (D) of a neutralizing anti-IL-1 mAb. Note the significantly reduced angio- genic response in D, similar to that observed in the CAM treated with PBS (E). F and G, IL-1RI expression in the CAM. RT-PCR was performed using specific chicken IL-1RI primers on retrotranscribed CAM mRNA (RT). Nonretrotranscribed CAM mRNA ( RT) was used as control (F). Paraffin-embedded CAM sections were nuclear counterstained and deco- rated with affinity-purified goat polyclonal anti-IL-1RI Ab on day 12. IL- 1RI immunoreactivity was evident in the endothelium lining the blood vessels (G). Original magnification, 400. No specific signal was observed in CAM in which primary Ab was replaced by preimmune rabbit serum (data not shown).

Article Snippet: Immunohistochemistry and Western blot analysis IL-1RI immunodetection was performed on deparaffinized 5- m CAM sections (15), using a goat polyclonal anti-mouse IL-1RI Ab (R&D Systems) and a preimmune goat serum as negative control.

Techniques: Chemotaxis Assay, Control, Boyden Chamber Assay, Expressing, Reverse Transcription Polymerase Chain Reaction