Journal: Current Issues in Molecular Biology

Article Title: Transcriptomic Establishment of Pig Macrophage Polarization Signatures

doi: 10.3390/cimb45030151

Figure Lengend Snippet: Brief methods of bone marrow-derived macrophage induction.

Article Snippet: M2_IL4 + IL10 , M2 , 10 ng/mL IL10(Proteintech, USA, HZ-1145), and 10 ng/mL IL4 (Proteintech, Rosemount, IL, USA, HZ-1004).

Techniques:

Journal: Current Issues in Molecular Biology

Article Title: Transcriptomic Establishment of Pig Macrophage Polarization Signatures

doi: 10.3390/cimb45030151

Figure Lengend Snippet: Transcriptomic comparison of porcine macrophage phenotypes, M1 (M1_IFNγ + LPS, and M1_GM-CSF) versus M2 (M1_IFNγ + LPS, and M1_GM-CSF). ( A ) PCA clustering M1_IFNγ + LPS ( n = 4), M1_GM-CSF ( n = 4), M2_IL4 + IL10 ( n = 3), M2_M-CSF ( n = 3); ( B ) Top enriched GO biological processes of 730 DEG genes, M1 (M1_IFNγ + LPS, and M1_GM-CSF) and M2 (M2_IL4 + IL10, and M2_M-CSF); ( C ) Heatmap exhibiting 49 classical macrophage marker genes expression profile among M1_IFNγ + LPS, M1_GM-CSF, M2_IL4 + IL10, and M2_M-CSF; ( D ) The DEGs between M1 and M2 were studied by using the String online tool. The interaction between each protein pair is represented by lines, and the size of the circle is directly proportional to the degree.

Article Snippet: M2_IL4 + IL10 , M2 , 10 ng/mL IL10(Proteintech, USA, HZ-1145), and 10 ng/mL IL4 (Proteintech, Rosemount, IL, USA, HZ-1004).

Techniques: Comparison, Marker, Expressing

Journal: Current Issues in Molecular Biology

Article Title: Transcriptomic Establishment of Pig Macrophage Polarization Signatures

doi: 10.3390/cimb45030151

Figure Lengend Snippet: Comparison of transcriptomics in different phenotypes of porcine macrophages. ( A ) Heatmap showing that specific macrophage phenotypes were aggregated by the induction method, M1_IFNγ + LPS, and M1_GM-CSF; ( B ) Heatmap showing that specific macrophage phenotypes were aggregated by the induction method M2_IL4 + IL10 and M2_M-CSF; ( C ) GO pathway enrichment analysis of two polarized macrophages, M1_IFNγ + LPS and M1_GM-CSF; ( D ) GO pathway enrichment analysis of two polarized macrophages, M2_IL4 + IL10 and M2_M-CSF; ( E ) The DEGs were studied by using the String online tool, M1_IFNγ + LPS, and M1_GM-CSF. The interaction between each protein pair is represented by lines, and the size of the circle is directly proportional to the degree of interaction. We adopt a node number greater than 1; ( F ) The DEGs are studied by using the String online tool, M2_IL4 + IL10, and M2_M-CSF. The interaction between each protein pair is represented by lines, and the size of the circle is directly proportional to the degree of interaction. We adopt a node number greater than 2.

Article Snippet: M2_IL4 + IL10 , M2 , 10 ng/mL IL10(Proteintech, USA, HZ-1145), and 10 ng/mL IL4 (Proteintech, Rosemount, IL, USA, HZ-1004).

Techniques: Comparison

Journal: Current Issues in Molecular Biology

Article Title: Transcriptomic Establishment of Pig Macrophage Polarization Signatures

doi: 10.3390/cimb45030151

Figure Lengend Snippet: Validation of M1_IFNγ + LPS, M1_GMCSF, M2_M-CSF, and M2_IL4 + IL10 signatures. ( A ) SS2-infected macrophages were enriched in M1_IFNγ + LPS based on GSEA (Data source: Microarray analysis); ( B ) PRRSV-infected macrophages were enriched in M1_GM-CSF based on GSEA (Data source: RNA-seq); ( C ). T. gondii -infected macrophages were enriched in M2_IL4 + IL10 based on GSEA (Data source: RNA-seq); ( D ) T. gondii Me49 -infected macrophages were enriched in M2_M-CSF based on GSEA (Data source: RNA-seq).

Article Snippet: M2_IL4 + IL10 , M2 , 10 ng/mL IL10(Proteintech, USA, HZ-1145), and 10 ng/mL IL4 (Proteintech, Rosemount, IL, USA, HZ-1004).

Techniques: Biomarker Discovery, Infection, Microarray, RNA Sequencing

Journal: ACS Applied Materials & Interfaces

Article Title: IL-4/Nanohydroxyapatite Codelivery in a Dual-Bionic Scaffold: Design and Immunomodulatory Endogenous Osteogenesis Mechanism

doi: 10.1021/acsami.5c23172

Figure Lengend Snippet: Schematic illustration of the bioactive PHS/SG@IL-4 dual-bionic scaffold: from bionic design to critical-sized calvarial regeneration. Inspired by the “soft-hard” integration structure of natural bone, we fabricated PCL/nHA composite filaments to construct gyroid bionic scaffolds through FDM 3D printing. Subsequently, SG hydrogel loaded with interleukin-4 (SG@IL-4) was infused into the scaffold pores using a vacuum-assisted technique. The resulting scaffold provides sustained release of IL-4 and bioactive nHA. These signals work synergistically to polarize macrophages toward the M2 phenotype and subsequently promote bone regeneration by modulating the PI3K-AKT and calcium signaling pathways.

Article Snippet: The SG/IL-4 hydrogel was prepared by homogeneously mixing the SG hydrogel precursor solution with recombinant mouse interleukin-4 (IL-4, Solarbio, China, cat. no. P00196) to achieve a final concentration of 1 μg/mL.

Techniques: Construct, Protein-Protein interactions

Journal: Veterinary immunology and immunopathology

Article Title: Interleukin-4 (IL-4) may regulate alternative activation of macrophage-like cells in chickens: A sequential study using novel and specific neutralizing monoclonal antibodies against chicken IL-4.

doi: 10.1016/j.vetimm.2018.10.011

Figure Lengend Snippet: Fig. 1. Detection of recombinant yeast chIL-4 by western blot analysis. (A) Western blot detection of yeast chIL-4 using mouse anti-chIL-4 monoclonal antibody #6D9G and cross reactivity of #6D9G with mouse and human IL-4 as well as chicken IL-13, IL-10 and IFN-γ, (B) Blot showing detection of chIL-4 using mouse anti- chIL-4 monoclonal antibody #8GE10 and cross reactivity of ##8GE10 with mouse and human IL-4 as well as chicken IL-13, IL-10 and IFN-γ, (C) chIL-4 detection using rabbit polyclonal antibody #53 against chIL-4. Molecular weight (kDa) marker is presented on the left. Data represent three independent experiments.

Article Snippet: Similarly, the cross reactivity of these mAbs was determined using mouse and human IL-4 as well as chicken chIL-13, chIL-10 and chIFN-γ at a concentration of 1 μg/lane (all from Kingfisher biotech, St. Paul, MN, USA).

Techniques: Recombinant, Western Blot, Molecular Weight, Marker

Journal: Nature Communications

Article Title: CD163 interacts with TWEAK to regulate tissue regeneration after ischaemic injury

doi: 10.1038/ncomms8792

Figure Lengend Snippet: ( a ) TWEAK (100 ng ml −1 ) was added to VEGFR1, Fn14 or CD163-coated wells. Bound protein was measured by ELISA using an antibody to detect TWEAK. ( b ) Fn14 or CD163 were coated onto 96-well plates and incubated with increasing doses of untagged TWEAK. Bound protein was measured by ELISA. ( c ) Immunoblotting of MDEC stimulated with TWEAK at 200 ng ml −1 (which activates both canonical and non-canonical NF-κB signalling) and increasing doses of recombinant mouse sCD163 for the indicated proteins. ( d ) Immunoprecipitation of serum before and after femoral ligation using anti-TWEAK antibody with immunoblotting for CD163. (The figures show a representative blot with a total of four experiments conducted.) ( e ) Immunostaining for VE-cadherin and β-dystroglycan (red) in limbs of CD163 −/− mice treated for 14 days with saline (control), TWEAK administration (25 μg kg −1 day −1 administered via Alzet pump) alone or in combination with sCD163 administration (25 μg kg −1 day −1 administered by intraperitoneal injection) ( n =5 per group). The graphs show the number of VE-cadherin positive cells per myofibre and the average muscle fibre area per group. ( f ) Immunblotting of CD163 −/− limb after 14 days of TWEAK administration (25 μg kg −1 day −1 administered via Alzet pump) alone or in combination with sCD163 administration (25 μg kg −1 day −1 administered by intraperitoneal injection) ( n =5 per group). O.D., optical density. Bar graphs show quantitation of densitometry for phosphorylated to total p65, p52 and NICD. * P <0.05 versus other groups. For multiple group comparisons, we utilized a one-way ANOVA. If the variance ratio test ( F -test) was significant, a more detailed post hoc analysis of differences between groups was made using a Tukey–Kramer honest significance difference test. For binding experiments, average of at least four well per group shown with each experiment repeated at least two times with representative results shown.

Article Snippet: For experiments shown in , mouse recombinant TWEAK (untagged) was obtained from Kingfisher Biotech.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Western Blot, Recombinant, Immunoprecipitation, Ligation, Immunostaining, Saline, Control, Injection, Quantitation Assay, Binding Assay