tlr4  (MedChemExpress)

Journal: Frontiers in Immunology

Article Title: Adsorption-mediated modulation of lipopolysaccharide bioactivity by clinoptilolite zeolite with in vitro immunomodulatory effects and in vivo safety evaluation

doi: 10.3389/fimmu.2026.1818740

Figure Lengend Snippet: TLR4 inhibition supports zeolite-mediated reduction of LPS-induced NF-κB activation in THP-1 Dual cells. THP-1 Dual NF-κB reporter cells were pre-treated with the small-molecule TLR4 inhibitor TAK-242 (5 µM) for 1 h prior to stimulation with lipopolysaccharide (LPS) or combined clinoptilolite zeolite and LPS treatment conditions. DMSO (0.1%) was included as a vehicle control. Data are presented as mean ± SD. Statistical significance was determined using an unpaired Student’s t-test. ns, not significant.

Article Snippet: To inhibit TLR4 signaling, cells were pre-treated with the small-molecule TLR4 inhibitor TAK-242 ( ) (MedChemExpress, HY-11109) at a final concentration of 5 μM for 1 h prior to stimulation.

Techniques: Inhibition, Activation Assay, Control

Journal: Advanced Science

Article Title: Integrated Single‐Cell and TCR Profiling Reveals Protection‐Associated CD8 + T Cell Subsets Linked to Viral Control in PRRSV

doi: 10.1002/advs.76732

Figure Lengend Snippet: CD8 + T cells preferentially receive signals from macrophages /monocytes for activation via TLR4 signaling. (A) Communications between APC and CD8 + T cells within each experimental group as inferred by CellPhoneDB. (B) Analysis of the expression level of antigen presentation‐related genes in indicated cell subclusters. (C) Dot plot showing normalized mean expression level of genes involved in innate immunity in indicated subclusters of macrophages and monocytes. (D) The KEGG enrichment analysis of innate immunity‐associated pathways for differentially upregulated genes in the subclusters of macrophages and monocytes. The enrichment level is determined by the number of included genes and their expression levels. (E) Percentage of cells containing virus RNA fragments in the indicated cell clusters. (F, G) Role of TLR4 signaling in T cell activation. The spleen cell mixture from three pigs of JXA1R groups at 5dpc were prepared. The ability to secret IFN‐γ was determined using ELISpot in the presence of TAK‐242, a TLR4 inhibitor or absence of macrophage/monocytes that were depleted by mouse monoclonal antibodies to CD14. The representative ELISpot wells (F) and statistical analysis (G) were presented. (H, I) Role of TLR8 signaling in T cell activation with TLR8 inhibitor. The representative ELISpot wells (H) and statistical analysis (I) were presented. Statistical analysis was performed by two‐tailed Student's t‐test, and error bars indicate means ± standard error of mean (SEM). Asterisks (*) indicate the statistical significance: *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: For the TLR‐4 or TLR‐8 loss‐function assay, the CD14‐deleted cells were pre‐incubated with TAK‐242 (MCE, #243984‐11‐4) or CU‐CPT8m (MCE, #HY‐112050) at a final concentration of 5 μM for 2 h, followed by the addition of virus, and ELISpot was then performed.

Techniques: Activation Assay, Expressing, Immunopeptidomics, Virus, Enzyme-linked Immunospot, Bioprocessing, Two Tailed Test

Journal: Advanced Science

Article Title: Integrated Single‐Cell and TCR Profiling Reveals Protection‐Associated CD8 + T Cell Subsets Linked to Viral Control in PRRSV

doi: 10.1002/advs.76732

Figure Lengend Snippet: Schematic model summarizing immune features associated with protection and pathogenesis. In the protected group, antigen presentation is associated with specialized subset of macrophages and monocytes characterized by high TLR4 and pro‐inflammatory signals expression. With appropriate CD4 + T cell help, these CD8 + T cells acquire robust cytotoxic effector functions and mediate efficient antiviral immunity. In contrast, in unprotected animals, antigen presentation is associated with monocyte populations expressing suppressive and inhibitory signals. CD8 + T cells in this context exhibit increased expression of exhaustion‐associated markers, especially CTLA4 , together with reduced effector functionality. Aberrant CD4 + T cell–CD8 + T cell communication further reinforces T cell exhaustion, ultimately resulting in impaired antiviral responses and disease progression.

Article Snippet: For the TLR‐4 or TLR‐8 loss‐function assay, the CD14‐deleted cells were pre‐incubated with TAK‐242 (MCE, #243984‐11‐4) or CU‐CPT8m (MCE, #HY‐112050) at a final concentration of 5 μM for 2 h, followed by the addition of virus, and ELISpot was then performed.

Techniques: Immunopeptidomics, Expressing, Biomarker Discovery

Journal: Molecular Biomedicine

Article Title: Oncolytic virus OHSV2 induces pyroptosis in bladder cancer cells via the TLR4/NLRP3/Caspase-1/GSDMD pathway

doi: 10.1186/s43556-026-00506-4

Figure Lengend Snippet: OHSV2 induces NLRP3/Caspase-1/GSDMD-dependent pyroptosis in bladder cancer cells via TLR4. a RT-qPCR analysis of the expression levels of TLR4 in bladder cancer cells treated with different concentrations (5% PBS, MOI = 1, MOI = 5) (n = 3). b Western Blot analysis of the expression levels of TLR4 in bladder cancer cells treated with different concentrations (5% PBS, MOI = 1, MOI = 5). c Immunohistochemistry analysis to detect the expression of TLR4 proteins in subcutaneous xenograft tumor tissues from nude mice. d CCK-8 assay to evaluate the effects of OHSV2 in combination with an TLR4 inhibitor (Resatorvid) on the viability of bladder cancer cells (n = 6). e Western Blot analysis to evaluate the effects of OHSV2 in combination with an TLR4 inhibitor (Resatorvid) on the expression of TLR4, NLRP3, Caspase-1, and GSDMD. f Western blot analysis of TLR4 after TLR4 knockdown. g CCK-8 assay showing partial restoration of cell viability after TLR4 knockdown (n = 6). h Western blot analysis of NLRP3, Cleaved Caspase-1, and N-GSDMD expression in TLR4-overexpressing cells after OHSV2 treatment. i Western blot analysis of NLRP3, Cleaved Caspase-1, and N-GSDMD expression in TLR4-knockdown cells after OHSV2 treatment. Data are presented as mean ± SD. Statistical significance was determined by unpaired two-tailed Student's t-test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: The TLR4 agonist RS09 (MCE, USA) was administered intraperitoneally at a dose of 2 mg/kg per injection every day from day 0.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemistry, CCK-8 Assay, Knockdown, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Oral Sodium Hyaluronate Reshapes Gut Microbiota Composition and Suppresses the LPS-TLR4/NF-κB Pathway to Exert Neuroprotection in MPTP-Induced Parkinson’s Disease

doi: 10.3390/ijms27156573

Figure Lengend Snippet: Oral SH inhibited the TLR4/MyD88/NF-κB signaling pathways both in the SN and the colon of MPTP-induced mice. ( A ) Schematic representation of the signaling pathway. ( B ) Representative WB bands of TLR4, MyD88, and NF-κB in the colon. ( C ) Representative WB bands of TLR4, MyD88, and NF-κB in the midbrain containing the SN. ( D – F ) The density analysis results of TLR4, MyD88, and NF-κB in the colon. ( G – I ) The density analysis results of TLR4, MyD88, and NF-κB in the midbrain containing the SN. ( n = 3 for each group. Data are presented as mean ± SD. *** p < 0.001 versus the control group; ### p < 0.001 versus the MPTP group.).

Article Snippet: Membranes were incubated at 4 °C overnight with the following primary antibodies: rabbit anti-β-actin antibody (1:50,000, AC026, Abclonal, Wuhan, China), mouse anti-TH antibody (1:1000, A0028, Abclonal, Wuhan, China), rabbit anti-TNF-α antibody (1:1000, A20851, Abclonal, Wuhan, China), rabbit anti-IL-1β antibody (1:1000, A11369, Abclonal, Wuhan, China), rabbit anti-IL-6 antibody (1:1000, A0286, Abclonal, Wuhan, China), goat anti-COX-2 antibody (1:1000, A1253, Abclonal, Wuhan, China), mouse anti-TLR4 antibody (1:1000, A5258, Abclonal, Wuhan, China), rabbit anti-MyD88 antibody (1:400, A0980, Abclonal, Wuhan, China), rabbit anti-IκB-α antibody (1:1000, A24742, Abclonal, Wuhan, China), rabbit anti-ZO-1 antibody (1:1000, A0659, Abclonal, Wuhan, China), mouse anti-claudin-1 antibody (1:1000, Abclonal, Wuhan, China), rabbit anti-occludin antibody (1:1000, Abclonal, China), and mouse anti-claudin-5 antibody (1:1000, A10207 , Abclonal, Wuhan, China).

Techniques: Protein-Protein interactions, Control