tlr2 inhibitor tl2 c29  (InvivoGen)

Journal: Journal of Inflammation Research

Article Title: Nasal Streptococcus pyogenes Infection Exacerbates Psoriasiform Relapse in Mice Through TLR2-Dependent Dendritic Cell Metabolic Reprogramming and γδ T Cell Activation

doi: 10.2147/JIR.S596821

Figure Lengend Snippet: Streptococcal LTA promotes TLR2-dependent metabolic reprogramming and hyper-migration in BMDCs. ( A ) qPCR analysis of pro-inflammatory, metabolic (HK2, LDHA), and motility-related (MYPT1) genes in BMDCs stimulated with LTA (100 ng/mL) ± TLR2 inhibitor (TL2-C29). ( B ) Real-time metabolic profiles showing OCR and ECAR. ( C ) Representative images and quantification of the scratch wound healing assay. Red dashed lines indicate the wound edge at the endpoint (24 h), marking the position to which cells have migrated. Data are mean ± SEM; error bars indicate standard error of the mean. Statistical significance was determined by one-way ANOVA. Data are representative of five independent experiments; * P < 0.05, ** P < 0.01, **** P < 0.0001 vs Ctrl; # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 vs LTA.

Article Snippet: Stimulation: BMDCs were pretreated with 100 ng/mL purified S. pyogenes lipoteichoic acid (LTA; Merk, NO. L3140-5MG) for 6 hours as, with or without an incubation with the TLR2 inhibitor TL2-C29 (25 μM; InvivoGen, Cat.code: inh-c29).

Techniques: Migration, Wound Healing Assay

Journal: Journal of Inflammation Research

Article Title: Nasal Streptococcus pyogenes Infection Exacerbates Psoriasiform Relapse in Mice Through TLR2-Dependent Dendritic Cell Metabolic Reprogramming and γδ T Cell Activation

doi: 10.2147/JIR.S596821

Figure Lengend Snippet: TLR2 ligation activates the canonical NF-κB pathway and actomyosin contractility in DCs. Representative Western blots and densitometric quantification of p-P65, p-IKBα, NOS2, TLR2, p-MYPT1, and p-MLC in BMDCs following LTA treatment ± TL2-C29. Tubulin or β-actin served as the loading control. Data are mean ± SEM from n = 3 independent experiments. Statistical significance was determined by one-way ANOVA; * P < 0.05, ** P < 0.01vs Ctrl; # P < 0.05, ## P < 0.01vs LTA.

Article Snippet: Stimulation: BMDCs were pretreated with 100 ng/mL purified S. pyogenes lipoteichoic acid (LTA; Merk, NO. L3140-5MG) for 6 hours as, with or without an incubation with the TLR2 inhibitor TL2-C29 (25 μM; InvivoGen, Cat.code: inh-c29).

Techniques: Ligation, Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: Nasal Streptococcus pyogenes Infection Exacerbates Psoriasiform Relapse in Mice Through TLR2-Dependent Dendritic Cell Metabolic Reprogramming and γδ T Cell Activation

doi: 10.2147/JIR.S596821

Figure Lengend Snippet: Mechanistic model of infection-triggered psoriasis relapse. Schematic representation of the “nasal-skin” immune axis: Nasal SP infection → TLR2-driven DC glycolytic reprogramming → DC migration to skin → Conjugation with γδ T cells → IL-17A-driven relapse. Blue arrows indicate downregulation, and Orange arrows indicate upregulation following SP intervention.

Article Snippet: Stimulation: BMDCs were pretreated with 100 ng/mL purified S. pyogenes lipoteichoic acid (LTA; Merk, NO. L3140-5MG) for 6 hours as, with or without an incubation with the TLR2 inhibitor TL2-C29 (25 μM; InvivoGen, Cat.code: inh-c29).

Techniques: Infection, Migration, Conjugation Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Hemoglobin inhibits fibroblast-to-cardiomyocyte reprogramming via TLR2/TLR4-dependent chromatin compaction

doi: 10.1016/j.omtn.2026.102900

Figure Lengend Snippet: Hemoglobin mediates gene repression through TLR2 and TLR4 (A and B) A study was conducted to determine the effect of hemoglobin (Hb) on (A) fibroblast-to-cardiomyocyte reprogramming and (B) fibroblast gene expression. With respect to fibroblast-to-cardiomyocyte reprogramming, cardiac fibroblasts were transfected with either miR combo or a non-targeting control miR. 24 h later, the cells were incubated with vehicle, a TLR2 pharmacological inhibitor, a TLR4 pharmacological inhibitor, or a combination of both pharmacological inhibitors for 3 h. After incubation with the indicated pharmacological inhibitors, Hb was added (5 mg/mL) to the media. All media was replaced the next day. Fourteen days after miR transfection, cells were analyzed for expression of the indicated cardiomyocyte specific genes by qPCR. Expression values were normalized to the housekeeping gene Gapdh. N = 6–10. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant). t test was used to determine the significance between the miR combo groups (## p < 0.01, # p < 0.05; ns, not significant). With respect to fibroblast gene expression, cardiac fibroblasts were incubated with vehicle, a TLR2 pharmacological inhibitor, a TLR4 pharmacological inhibitor, or a combination of both pharmacological inhibitors for 3 h. After incubation with the indicated pharmacological inhibitors, Hb was added (5 mg/mL) to the media. All media was replaced the next day. Expression of the indicated fibroblast-specific genes was determined by qPCR and normalized to the housekeeping gene Gapdh. N = 10. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant).

Article Snippet: Hb (Millipore Sigma, H2625), TLR2 signaling inhibitor-TL2-C29 (InvivoGen, catalog no. inh-c29), and TLR4 inhibitor-CLI-095 (InvivoGen, catalog no. tlrl-cli95-4) were used.

Techniques: Gene Expression, Transfection, Control, Incubation, Expressing

Journal: Nature Communications

Article Title: A tissue-intrinsic mechanism sensitizes HIV-1 particles for TLR-triggered innate immune responses

doi: 10.1038/s41467-026-72586-3

Figure Lengend Snippet: a Schematic representation of PRRs and their cognate ligands. PRRs: green, adapter proteins: pink, PAMPs: red, transcription factors: blue. b PRR inhibitor screening. MDMs were challenged with S or DC HIV-1 NL4.3 R5 virions in presence or absence of PRR inhibitors. IL-6 release was measured by ELISA from MDM supernatants. Data is normalized to mock infected condition (gray dotted line). c Measurement of IL-6 release from the supernatants of MDMs challenged with S or DC HIV-1 virions in presence or absence of the indicated inhibitors. d Volcano plot illustrating the differential gene expression between MDMs challenged with DC virus and mock challenged cells for 6 h. The number of down- and upregulated genes are indicated. e Volcano plot as in d. with highlighted gene signatures. Genes induced by TLR2 alone (blue), TLR8 alone (brown), co-regulated (light green) or associated with “Innate Immune Response” genes (red). f Volcano plot illustrating the differential gene expression between MDMs challenged with suspension or collagen primed HIV-1 virus for 6 h. The number of down- and upregulated genes are indicated. g Volcano plot as in f. with highlighted gene signatures. Genes induced by TLR2 alone (blue), TLR8 alone (brown), co-regulated (light green) or associated with Innate Immune Response genes (red). Top deregulated genes belonging to different gene ontology terms are highlighted. h Gene ontology analysis of biological processes repressed or induced in MDMs challenged with collagen primed virions as compared to cells challenged with suspension virus for 6 h. FDR is indicated (only upregulated pathways have a q -value < 0.05). Top deregulated genes for each pathway are highlighted. Results represent the mean ± SD of 3 independent donors. Symbols indicate individual donors ( b , c ) or individual genes ( d–g ). Significance is indicated by p-values, and was calculated by two-tailed paired t-tests or Wilcoxon matched-pairs signed rank test ( b , c ), differential gene expression analysis was performed using DESeq2: genes with adjusted p-value < 0.05 were considered significant ( d–g ), for GSEA analysis, p -values were corrected using Benjamini-Hochberg FDR ( h ). Source data are provided as a file.

Article Snippet: 5 μM of cGAS inhibitor (G140, Invivogen), 8 μM of TLR 1/2 inhibitor (Cu-CPT22, Selleckchem), 49 μM of TLRs 4& 2/6 inhibitor (GIT27, Tocris), 2 μM of TLR4 inhibitor (CLI-095, Invivogen), 100 μM of TLR2 inhibitor (TL2-C29, Invivogen) or 10 μM of TLR8 inhibitor (Cu-CPT9a, Invivogen) were incubated with MDMs 3 h prior to infection.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Gene Expression, Virus, Suspension, Two Tailed Test

Journal: Nature Communications

Article Title: A tissue-intrinsic mechanism sensitizes HIV-1 particles for TLR-triggered innate immune responses

doi: 10.1038/s41467-026-72586-3

Figure Lengend Snippet: a Representative structure of an HIV-1 Env trimer. Epitopes targeted by different bNAbs or nNAbs as well as sCD4 are highlighted (based upon a prediction of NL4.3 Env trimer structure using AlphaFold 3). b Experimental workflow. HIV-1 NL4.3 R5 Vpr.Int.GFP virions harvested from suspension or collagen cultures were sequentially incubated with primary, epitope-specific anti-gp120 antibodies, then with AF568 coupled secondary antibodies prior to processing for microscopy analysis. c . Representative micrographs of stained virions. Vpr.Int.GFP positive spots (green) can be seen bound by different antibodies or sCD4-PE (red). Scale bar: 0.5 µm. d Spider plot representing the binding frequency of antibody binding to HIV-1 NL4.3 R5 as in (c). e . sTLR2 is retained at the surface of NL4.3 R5 but not NL4.3 ΔEnv virions after collagen priming. One representative Western Blot analysis showing the ratio from HIV-1 NL4.3 virions cultured in S, DC or LC. The TLR2/p24 ratios are indicated below the blots after correction, after subtraction of the TLR2 background signals from the last two lanes. f Quantification of western blot analyses from three independent experiments as in e . g . Representative micrographs. MDMs challenged with HIV1 NL4.3 R5 Vpr.Int.GFP virions after culture in suspension or in DC. Yellow arrows indicate Vpr.Int.GFP/TLR8 colocalization. Scale bar: 5 µm. h Quantification of Virus/TLR8 colocalization from micrographs as in d. Results represent the mean ± SD of three independent experiments. Significance is indicated by p -values, and was calculated by two-tailed paired t-tests ( d , f , h ). Source data are provided as a file.

Article Snippet: 5 μM of cGAS inhibitor (G140, Invivogen), 8 μM of TLR 1/2 inhibitor (Cu-CPT22, Selleckchem), 49 μM of TLRs 4& 2/6 inhibitor (GIT27, Tocris), 2 μM of TLR4 inhibitor (CLI-095, Invivogen), 100 μM of TLR2 inhibitor (TL2-C29, Invivogen) or 10 μM of TLR8 inhibitor (Cu-CPT9a, Invivogen) were incubated with MDMs 3 h prior to infection.

Techniques: Suspension, Incubation, Microscopy, Staining, Binding Assay, Western Blot, Cell Culture, Virus, Two Tailed Test