Journal: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: (A) Normalized Kratky plot of IL-1RI-ECD (black), IL-1RII-ECD (red), ST2-ECD (blue) and IL-1RAcP-ECD (cyan). IL-1RII-ECD (red) showed a bell-shaped plot, indicating a structure that is less flexible. (B) Normalized pair distribution function P(r) of IL-1RI-ECD (black), IL-1RII-ECD (red), ST2-ECD (blue) and IL-1RAcP-ECD (cyan). The q-range used for P(r) calculations is as follows: IL-1 RI-ECD (0.01730.2758Å−1); IL-1RII-ECD (0.0142 to 0.2634 Å−1); ST2-ECD (0.0125 to 0.2770 Å−1); IL-1RAcP-ECD (0.0240 to 0.2946 Å−1). The dotted rectangles indicate the different features between IL-1 receptors. (C) Fitting of experimental data (black dots) to the theoretical profiles of the initial (blue), the best-fit (red) and ensemble model (green) of IL-1RI-ECD, IL-1RII-ECD, ST2-ECD and IL-1RAcP-ECD (up panel). Error-weighted residuals of the fitting after FFT smoothing (lines) (low panel). (D) The weighted Rg distribution of selected 100 two-state ensembles of IL-1RI-ECD, IL-1RII-ECD, ST2-ECD, and IL-1RAcP-ECD. The top two-state ensemble fitting the experimental data is shown as cartoon for visualization (glycans are shown as spheres). The peak height reflects the weight of the conformational state. (See also Figures S2 and S3, Table S1, Table S2 and Table S3)

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques:

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: (A) Schematic presentation of the binding affinity measurement. Ligand: IL-1RAcP-ECD or D2/D3 linker-replaced IL-1RAcP-ECD mutants (IL-1RAcP-ECD-RI, IL-1RAcP-ECD-RII, IL-1RAcP-ECD-ST2); Analyte: the preformed binary complex IL-33/ST2-ECD or IL-1β/IL-1RI-ECD. (B) The binding of wild-type or mutated IL-1RAcP-ECD to the preformed binary complexes IL-33/ST2-ECD (up panel) and IL-1β/IL-1 RI-ECD (low panel). The SPR sensorgrams (black line) were fitted to a 1:1 Langmuir binding model (colored lines). (C) Comparison of the binding affinity of wild-type and D2/D3 linker-replaced IL-1RAcP-ECD. On rate (kon), off rate (koff) and dissociation constant (KD) for the binding of IL-1RAcP-ECD and IL-1RAcP-ECD mutants to IL-33/ST2-ECD or IL-1β/IL-1RI-ECD were summarized. (See also Figure S1 and S5A)

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques: Binding Assay

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: The signal of NF-κB activation was measured in 293T-IL-1RAcP-KO cells, which were stimulated for 6h with IL-1 cytokines, including IL-1β (A), IL-33 (B), and IL-36γ (C) with the co-transfection of the respective primary receptors, IL-1RAcP or D2/D3 linker-replaced IL-1RAcP, and luciferase reporter genes. For IL-1β-mediated NF-κB signaling, only IL-1RAcP was transfected because of endogenous expression of IL-1Rl. The presented results are the means αS.D. from triplicate experiments. The protein levels of various IL-1RAcP were detected by western blot analysis with the whole cell lysate. The experiments were performed independently at least three times. (See also Figures S4 and S5C)

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques: Activation Assay, Cotransfection, Luciferase, Transfection, Expressing, Western Blot

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: The signal of NF-κB activation was measured in 293T-IL-1RAcP-KO cells upon stimulation with IL-1β (A), IL-33 (B) and IL-36γ (C), by co-transfection of primary receptors with IL-1RAcP, IL-1 RAcPK238A, IL-1 RAcPD239A or IL-1RAcPK238A/D239A using the dual luciferase reporter assay. The presented results are the means αS.D.from triplicate experiments. The consistency of protein levels of mutated IL-1RAcP was confirmed by western blot analysis with the whole cell lysis. The experiments were performed independently at least three times. (See also Figures S4, S5C and S6)

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques: Activation Assay, Cotransfection, Luciferase, Reporter Assay, Western Blot, Lysis

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: (A) Fitting of the experimental data (black dots) to the theoretical profiles of the initial (blue), the best-fit (red) and ensemble model (green) of D2/D3 linker replaced IL-1RAcP mutants. (B) Error-weighted residuals of the fitting after FFT smoothing (lines). (C) The weighted Rg distribution of selected 100 two-state ensembles of D2/D3 linker-replaced IL-1 RAcP-ECD mutants. The top two-state ensemble fitting the experimental data is shown as cartoon for visualization (glycans are shown as spheres). The D1D2 module was in the same orientation as that of wild-type IL-1RAcP-ECD, while the D3 domain was rotated in a different orientation. The peak height reflects the weight of the conformational state and reveals the difference between wild-type IL-1RAcP-ECD and the mutants. (See also Figures S2 and S3, Table S1, Table S2 and Table S3)

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques:

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques: Recombinant, Plasmid Preparation, Software

Journal: PLoS ONE

Article Title: PPAR Agonist-Induced Reduction of Mcp1 in Atherosclerotic Plaques of Obese, Insulin-Resistant Mice Depends on Adiponectin-Induced Irak3 Expression

doi: 10.1371/journal.pone.0062253

Figure Lengend Snippet: Primers used for qRT-PCR analysis.

Article Snippet: To detect protein levels, membranes were incubated with primary antibodies against β-actin (Cell Signaling Technology) and Irak3 (Rockland Immunochemicals).

Techniques:

Journal: PLoS ONE

Article Title: PPAR Agonist-Induced Reduction of Mcp1 in Atherosclerotic Plaques of Obese, Insulin-Resistant Mice Depends on Adiponectin-Induced Irak3 Expression

doi: 10.1371/journal.pone.0062253

Figure Lengend Snippet: ( A ) Representative Mac-3 staining of aortic sinus plaques of placebo-, fenofibrate- and rosiglitazone-treated DKO mice at 24 weeks. ( B ) Gene expression in the aorta was analyzed by measuring relative RNA levels using qRT-PCR for Tnfα , IL6 , Mcp1 and Irak3 . Data are means ± SEM. Scale bar = 500 µm. * P <0.05, ** P <0.01 and *** P <0.001 DKO compared with C57BL/6 J mice; $$ P <0.01 and $$$ P <0.001 PPAR agonist-treated compared with placebo-treated DKO mice; £ P <0.05, ££ P <0.01 and £££ P <0.001 rosiglitazone-treated compared with fenofibrate-treated DKO mice.

Article Snippet: To detect protein levels, membranes were incubated with primary antibodies against β-actin (Cell Signaling Technology) and Irak3 (Rockland Immunochemicals).

Techniques: Staining, Gene Expression, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: PPAR Agonist-Induced Reduction of Mcp1 in Atherosclerotic Plaques of Obese, Insulin-Resistant Mice Depends on Adiponectin-Induced Irak3 Expression

doi: 10.1371/journal.pone.0062253

Figure Lengend Snippet: ( A ) Soluble Mcp1 protein levels in DKO BMDM exposed to 50 µM fenofibrate, 10 µM rosiglitazone or 5 µM GW9662 for 24 hours as determined by ELISA. Data are means ± SEM; n = 16 from three different mice. $$ P <0.01 compared with fenofibrate-treated BMDM. ( B ) Irak3 RNA and protein levels of DKO BMDM exposed to 1 or 10 µg/mL globular adiponectin for 24 hours as determined by qRT-PCR and Western blotting. Data are means ± SEM; n = 6. *** P <0.001 compared with DKO BMDM; $ P <0.05 and $$ P <0.01 compared with DKO BMDM exposed to 1 µg/mL globular adiponectin. ( C ) Soluble Mcp1 protein levels (n = 18 from three different mice), NFκB p50 DNA binding activity (n = 8 from two different mice) and mROS production (n = 6) in IRAK3 −/− BMDM exposed to 50 µM fenofibrate or 10 µM rosiglitazone for 24 hours as determined by ELISA and flow cytometry. Data are means ± SEM. * P <0.05, ** P <0.01 and *** P <0.001 compared with C57BL/6 J BMDM; $ P <0.05 and $$$ P <0.001 compared with IRAK3 −/− BMDM; £££ P <0.001 compared with fenofibrate-treated BMDM. Abbreviations: BMDM, bone marrow-derived macrophages; mROS, mitochondrial reactive oxygen species.

Article Snippet: To detect protein levels, membranes were incubated with primary antibodies against β-actin (Cell Signaling Technology) and Irak3 (Rockland Immunochemicals).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Binding Assay, Activity Assay, Flow Cytometry, Derivative Assay

Journal: PLoS ONE

Article Title: PPAR Agonist-Induced Reduction of Mcp1 in Atherosclerotic Plaques of Obese, Insulin-Resistant Mice Depends on Adiponectin-Induced Irak3 Expression

doi: 10.1371/journal.pone.0062253

Figure Lengend Snippet: Plaque volume was determined by measuring lipid (oil red O)-stained surfaces in subsequent sections; macrophages were stained with anti-Mac-3 antibody. Gene expression in the aorta was analyzed by measuring relative RNA levels using qRT-PCR for Pparγ , Mcp1, Irak3 and Adipoq . Data are means ± SEM. ** P <0.01 and *** P <0.001 HFD-fed compared with SD-fed LDL-receptor deficient mice. Abbreviations: HFD, high fat diet; SD, standard diet.

Article Snippet: To detect protein levels, membranes were incubated with primary antibodies against β-actin (Cell Signaling Technology) and Irak3 (Rockland Immunochemicals).

Techniques: Staining, Gene Expression, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: PPAR Agonist-Induced Reduction of Mcp1 in Atherosclerotic Plaques of Obese, Insulin-Resistant Mice Depends on Adiponectin-Induced Irak3 Expression

doi: 10.1371/journal.pone.0062253

Figure Lengend Snippet: The schematic draw demonstrates the anti-atherosclerotic properties of the PPARγ agonist rosiglitazone. Treatment with rosiglitazone improves the adipocyte function characterized by a decrease in adipocyte size, a reduction in adipose tissue macrophages and an increased expression of anti-inflammatory adiponectin. The increase in blood adiponectin and de novo adiponectin production in atherosclerotic lesions is necessary for the upregulation of Irak3 in plaque macrophages, which is crucial for the indirect rosiglitazone-mediated decrease in Mcp1 secretion. Abbreviations: Mφ, macrophages; ROS, reactive oxygen species.

Article Snippet: To detect protein levels, membranes were incubated with primary antibodies against β-actin (Cell Signaling Technology) and Irak3 (Rockland Immunochemicals).

Techniques: Expressing