rabbit monoclonal anti tlr2  (Servicebio Inc)

Journal: Frontiers in Immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: Primer sequences used in quantitative PCR.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-κB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit anti-TNF-α polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1β monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-β-actin monoclonal antibody (1:2,000 dilution, Beyotime Biotechnology, China) were used as the primary antibodies in this study.

Techniques: Sequencing

Journal: Frontiers in Immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: Targeting of TLR2 3′-UTR by novel-miR1. (A) Venn diagram showing the intersection of predicted target genes from different software programs, namely, PITA, miRanda, and TargetScan. (B) Gene Ontology (GO) molecular function annotations of the target genes of novel-miR1. The top 10 GO terms for biological processes, cellular component, and molecular function are shown based on their respective p-values. (C) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the predicted target genes of novel-miR1. The size of dots indicates the number of genes associated with each pathway. (D) Overexpression of novel-miR1 in rabbit PBLCs. (E) Relative expression of novel-miR1 target genes in rabbit PBLCs after novel-miR1 treatment. (F) novel-miR1 binding TLR2 3′-UTR site and the sequences of luciferase reporter plasmids (pmir-GLO-TLR2-WT and pmir-GLO-TLR2-Mut). (G) Luciferase reporter assay results indicating the downregulation of TLR2 through binding to its target 3′-UTR. (H) Western blot assay results showing the downregulation of TLR2 protein in PBLCs by novel-miR1. (I) Western blot assay results demonstrating the downregulation of TLR2 protein in PBLCs by novel-miR1 (fold change was calculated by the gray values of the image). ns, p ≥ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-κB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit anti-TNF-α polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1β monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-β-actin monoclonal antibody (1:2,000 dilution, Beyotime Biotechnology, China) were used as the primary antibodies in this study.

Techniques: Software, Over Expression, Expressing, Binding Assay, Luciferase, Reporter Assay, Western Blot

Journal: Frontiers in Immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: Inhibition of TLR2 signaling pathway and NF-κB activation by novel-miR1. (A) qPCR results demonstrate the reduction of TLR2, TNF-α, IL-1β, and IL-6 mRNA levels by novel-miR1. Each test was performed in triplicate. Fold change was calculated based on qPCR results. (B) Western blot assay results indicate the decrease in TLR2, P65, pP65, TNF-α, IL-1β, and IL-6 protein levels by novel-miR1. (C) Western blot assay results showing the decrease in protein levels of TLR2, P65, pP65, TNF-α, IL-1β, IL-6 (fold change was calculated by the gray values of the image). ns, p ≥ 0.05; *, p ≤ 0.05; ***, p ≤ 0.001.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-κB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit anti-TNF-α polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1β monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-β-actin monoclonal antibody (1:2,000 dilution, Beyotime Biotechnology, China) were used as the primary antibodies in this study.

Techniques: Inhibition, Activation Assay, Western Blot

Journal: Frontiers in Immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: C. pisiformis exosomes inhibit the TLR2 signaling pathway and NF-κB activation. (A) C. pisiformis exosomes can enter rabbit PBLCs. (B) qPCR results demonstrate the increased expression of novel-miR1 and decreased TLR2 mRNA level in rabbit PBLCs after treatment with C. pisiformis exosomes. Each test was performed in triplicate; fold change was calculated based on qPCR results. (C) Western blot assay results suggest that treatment with C. pisiformis exosomes decreased TLR2, P65, and pP65 protein expression levels. (D) Western blot assay results indicating the reduction in protein expression levels of TLR2, P65, and pP65 by C. pisiformis exosomes (fold change was calculated by the gray values of the image). *, p ≤ 0.05; ***, p ≤ 0.001.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-κB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit anti-TNF-α polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1β monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-β-actin monoclonal antibody (1:2,000 dilution, Beyotime Biotechnology, China) were used as the primary antibodies in this study.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: C. pisiformis inhibits the TLR2 signaling pathway and NF-κB activation. (A) Expression profile of novel-miR1 in rabbit PBLCs over a 3-month period. (B) Relative expression TLR2 mRNA in rabbit PBLCs during the initial 3-month period. (C) Western blot assay results indicate inhibition of TLR2, P65, and pP65 protein levels in C. pisiformis -infected rabbit PBLCs. (D) Western blot assay results demonstrating the inhibition of TLR2, P65, and pP65 protein levels in C. pisiformis -infected rabbit PBLCs (the fold change was calculated by the gray values of the image). (E) ELISA assay results indicate inhibition of IL-6, TNF-α, and IL-1β protein expression levels in C. pisiformis -infected rabbit PBLCs. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-κB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit anti-TNF-α polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1β monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-β-actin monoclonal antibody (1:2,000 dilution, Beyotime Biotechnology, China) were used as the primary antibodies in this study.

Techniques: Activation Assay, Expressing, Western Blot, Inhibition, Infection, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: Graphical representation illustrating the mechanism of novel-miR1 targeting TLR2 in PBLCs to regulate the immune response in rabbits. The figure was created using Figdraw.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-κB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit anti-TNF-α polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1β monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-β-actin monoclonal antibody (1:2,000 dilution, Beyotime Biotechnology, China) were used as the primary antibodies in this study.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Calcium-Based Antimicrobial Peptide Compounds Attenuate DNFB-Induced Atopic Dermatitis-Like Skin Lesions via Th-Cells in BALB/c Mice

doi: 10.3390/ijms231911371

Figure Lengend Snippet: TLR2 and TLR4 immunohistochemical staining, protein expression in dorsal skin lesions, and qRT-PCR in each group of BALB/c mice. ( a ) Immunohistochemical staining of TLR2 in dorsal AD-like skin lesion (200×). ( b ) Immunohistochemical staining of TLR4 in dorsal AD-like skin lesion. ( c , d ) Expression of TLR2 was significantly increased in the DT, DVT, DCT, and DDT groups, in comparison to the NT group, and the expression was significantly decreased in the DCT and DDT groups, in comparison to the DT and DVT groups in skin lesions. ( e , f ) Expression of TLR4 was significantly increased in the DT, DVT, DCT, and DDT groups, in comparison to the NT group, and the expression was significantly decreased in the DCT and DDT groups, in comparison to the DT and DVT groups in skin lesions (data are presented as the mean ± S.E.M; ns: no significance, *** p < 0.001, and **** p < 0.0001).

Article Snippet: The rabbit monoclonal TLR2 antibody (ab209216) and the mouse monoclonal TLR4 antibody (ab22048) were provided by Abcam Trading Co. (Shanghai, China).

Techniques: Immunohistochemical staining, Staining, Expressing, Quantitative RT-PCR, Comparison

Journal: iScience

Article Title: Neuregulin-1/ErbB4 signaling modulates Plasmodium falciparum HRP2-induced damage to brain cortical organoids

doi: 10.1016/j.isci.2022.104407

Figure Lengend Snippet: HRP2 increases pro-inflammatory and Toll-like receptor (TLR) genes in cortical organoids. HRP2-induced inflammation-related genes were profiled Fold changes greater than 1.5 and p values < 0.05 calculated using Student’s t -test was considered to be a significant dysregulation. (A) A heatmap was generated showing up-and down-regulated genes. The effects of HRP2 on cortical organoids at day 50 in culture are shown by site (B), including cerebral cortex and glia, as well as by pathway (C), including TLR signaling. Further analysis revealed the relationship between upregulated genes and activation of TLR2 pathway (D). Western blot densitometry of organoid lysates (data represented as means ± SEM) confirms that HRP2 significantly induces TLR2 and TLR1 expression, whereas NRG1 reduces these effects (E, F). a: p < 0.05 compared to basal conditions; b: p < 0.05 compared to HRP2 treatment. One-way ANOVA and t -test were used. Please also see Figure S4 .

Article Snippet: Rabbit monoclonal anti-human TLR2 (clone D769Z) , Cell Signaling , Cat #12276T RRID: AB_2797867.

Techniques: Generated, Activation Assay, Western Blot, Expressing

Journal: iScience

Article Title: Neuregulin-1/ErbB4 signaling modulates Plasmodium falciparum HRP2-induced damage to brain cortical organoids

doi: 10.1016/j.isci.2022.104407

Figure Lengend Snippet: HRP2 uses TLR1 and TLR2 to increase CXCL10 expression and cell death in iPSC Cell proliferation, apoptosis, and expression of inflammatory markers were assessed in iPSC treated with HRP2 after TLR1 and TLR2 blockage. (A) Proliferation assay showed that IPSC growth was partially restored, whereas their apoptosis (B) and necrosis (C) were reduced to the levels of untreated controls. TLR1 and TLR2 blocking in HRP2-treated iPSC has an antiapoptotic and anti-inflammatory effect, as it is confirmed by IHC: the expression of cleaved caspase 3 (D, F) and CXCL10 (E, G) is reduced. Data represented as means ± SEM a: p < 0.05 compared to basal conditions; b: p < 0.05 compared to HRP2 treatment. Statistical significance between groups was determined using t -test and analysis of variance.

Article Snippet: Rabbit monoclonal anti-human TLR2 (clone D769Z) , Cell Signaling , Cat #12276T RRID: AB_2797867.

Techniques: Expressing, Proliferation Assay, Blocking Assay

Journal: iScience

Article Title: Neuregulin-1/ErbB4 signaling modulates Plasmodium falciparum HRP2-induced damage to brain cortical organoids

doi: 10.1016/j.isci.2022.104407

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-human TLR2 (clone D769Z) , Cell Signaling , Cat #12276T RRID: AB_2797867.

Techniques: Functional Assay, Recombinant, Plasmid Preparation, CCK-8 Assay, Bicinchoninic Acid Protein Assay, Isolation, Gene Expression, Western Blot, Software, Microscopy