tlr9  (MedChemExpress)

Journal: Journal of Extracellular Vesicles

Article Title: Extracellular Vesicles Derived From Streptococcus anginosus Aggravate Lupus Nephritis by Triggering TLR2‐MyD88‐NF‐κB Signalling in NK Cells

doi: 10.1002/jev2.70134

Figure Lengend Snippet: Reduced frequency yet heightened cytotoxicity of peripheral blood NK cells in SLE patients. (a) Representative flow cytometry plots depicting gating strategy for TBNK cell populations. (b) Quantitative analysis of TBNK subpopulations in SLE patients ( n = 54) versus healthy controls (HCs; n = 43). (c) Correlation of peripheral blood NK cell percentage and SLEDAI score in SLE patients. (d) Intracellular expression of Granzyme B and TNF‐α in primary human NK cells following stimulation with PMA (10 ng/mL), ionomycin (1 µg/mL) and brefeldin A (BFA; 1 µg/mL) for 12 h, analysed by flow cytometry. (e) Quantification of granzyme B and TNF‐α expression. (f) Serum concentration of IL‐6, IL‐17A and TNF‐α in the serum of HC and SLE ( n = 12). (g) The cytotoxicity of NK against K‐562 cells at varying effector: target cell ratios. NK cells were purified from PBMC. NK cells and target cells were co‐cultured with tumour cells for 4 h. (h) Correlation between NK cell percentage and SLEDAI score in SLE patients ( n = 12). (i) Correlation of NK cell cytotoxicity with Streptococcus anginosus abundance in SLE patients ( n = 12). (j) Surface expression of TLR2, TLR4, TLR6, TLR7 and TLR9 on human NK cells. (k) Quantitative analysis of TLR receptor mean fluorescence intensity (MFI) on NK cells ( n = 6). Data are shown as mean ± SD, with individual data points representing biological replicates (average of technical duplicates). Statistical comparisons were performed using one‐way ANOVA with Tukey post‐tests. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns indicates not significant.

Article Snippet: TLR2 inhibitor (C29, Cat: HY‐100461), TLR4 (Stepharine, HY‐N9347), TLR7 (Enpatoran, HY‐134581A), TLR9 (AT791, HY‐124603) and MyD88 inhibitor (MyD88‐IN‐1, Cat: HY‐149992) were obtained from MedChemExpress (MCE, USA).

Techniques: Flow Cytometry, Expressing, Concentration Assay, Purification, Cell Culture, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Rabies Virus Regulates Inflammatory Response in BV-2 Cells through Activation of Myd88 and NF-κB Signaling Pathways via TLR7

doi: 10.3390/ijms25179144

Figure Lengend Snippet: Infection with RABV strains exhibiting varying levels of virulence activating TLR7 in mouse brain tissue: ( A ) illustrative representations of IHC analysis of TLR7 in sections of mouse brains infected with the RABV strains SC16, HN10 and CVS-11; ( B ) integrated optical density of TLR7 in mouse brain tissue. Statistical evaluations were conducted utilizing ANOVA (* p < 0.05, ** p < 0.01).

Article Snippet: The TLR7 inhibitor (HY-124603, MCE, Princeton, NJ, USA), agonist (HY-117602, MCE, USA) and Myd88 inhibitor (HY-50937) were purchased from Med-Chem-Express.

Techniques: Infection

Journal: International Journal of Molecular Sciences

Article Title: Rabies Virus Regulates Inflammatory Response in BV-2 Cells through Activation of Myd88 and NF-κB Signaling Pathways via TLR7

doi: 10.3390/ijms25179144

Figure Lengend Snippet: RABV inducing proinflammatory chemokines in vitro via TLR7. ( A ) N2a and BV-2 cells exposed to CVS-11, and subsequent Western blot analysis conducted to assess the expression of TLR7. ( B ) Quantitative assessment of the comparative signal intensities of TLR7. ( C ) Different RABV strains at an MOI of 0.1, 0.5, 1 and 10 of infected BV-2 cells. Cell culture supernatants were collected and the expression of CCL2 was measured. ( D ) The expression of CXCL10 of BV-2 cells infected with RABV at an MOI of 0.1, 0.5, 1 and 10. ( E ) The expression of IL-6 of BV-2 cells infected with RABV at an MOI of 0.1, 0.5, 1 and 10. ( F ) BV-2 cells incubated with or without the TLR7 inhibitor (HY-124603) before stimulation with CVS-11. The expression of CCL2 in the supernatant was measured via ELISA. ( G ) The expression of CXCL10 in the supernatant incubated with the TLR7 inhibitor (HY-124603) before stimulation with CVS-11. ( H ) The expression of IL-6 in the supernatant incubated with the TLR7 inhibitor (HY-124603) before stimulation with CVS-11. Statistical evaluations were conducted utilizing ANOVA (** p < 0.01).

Article Snippet: The TLR7 inhibitor (HY-124603, MCE, Princeton, NJ, USA), agonist (HY-117602, MCE, USA) and Myd88 inhibitor (HY-50937) were purchased from Med-Chem-Express.

Techniques: In Vitro, Western Blot, Expressing, Infection, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Rabies Virus Regulates Inflammatory Response in BV-2 Cells through Activation of Myd88 and NF-κB Signaling Pathways via TLR7

doi: 10.3390/ijms25179144

Figure Lengend Snippet: RABV inducing the activation of the Myd88 pathway through TLR7. ( A ) The expression levels of TLR7, MyD88, IRAK4 and TRAF6 assessed via Western blotting. ( B ) BV-2 cells pre-treated for 2 h with the inhibitor (HY-124603) and agonist (HY-117602) of TLR7 before stimulation with CVS-11 for 6 h. TRL7, Myd88, IRAK4 and TRAF6 were analyzed via Western blotting. ( C ) Quantitative assessment of the relative expression levels of TLR7, Myd88, IRAK4 and TRAF6, normalized against β-actin. ( D ) Quantitative assessment of the relative expression levels of TLR7, Myd88, IRAK4 and TRAF6 following normalization against β-actin, in response to treatment with TLR7 inhibitors (HY-124603) and agonists (HY-117602). ( E ) The expression levels of viral P gene mRNA, quantified using qRT-PCR following treatment with the TLR7 inhibitor (HY-124603) and agonist (HY-117602). ( F ) BV-2 cells pre-treated for 2 h with the TLR7 inhibitor (HY-124603) and for 30 min with the Myd88 inhibitor (HY-50937). The supernatants were collected, and the concentration of CCL2 was quantified at 24 h. ( G ) The expression of CXCL10 was measured at 24 h. ( H ) The expression of IL-6 was measured at 24 h. Statistical evaluations were conducted utilizing ANOVA (* p < 0.05, ** p < 0.01).

Article Snippet: The TLR7 inhibitor (HY-124603, MCE, Princeton, NJ, USA), agonist (HY-117602, MCE, USA) and Myd88 inhibitor (HY-50937) were purchased from Med-Chem-Express.

Techniques: Activation Assay, Expressing, Western Blot, Quantitative RT-PCR, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Rabies Virus Regulates Inflammatory Response in BV-2 Cells through Activation of Myd88 and NF-κB Signaling Pathways via TLR7

doi: 10.3390/ijms25179144

Figure Lengend Snippet: RABV inducing the phosphorylation and nuclear translocation of NF-κB. ( A ) BV-2 cells pre-treated for 2 h with the inhibitor (HY-124603) and agonist (HY-117602) of TLR7 before stimulation with CVS-11 for 30 min. NF-κB p-p65, NF-κB p65 and IκBα analyzed via Western blotting. ( B ) Analysis of the nuclear translocation of NF-κB in BV-2 cells incubated with or without the TLR7 inhibitor (HY-124603) with RABV infection for 30 min, carried out using Image Stream. ( C ) BV-2 cells under 2 h pre-treatment with the TLR7 inhibitor (HY-124603) and 1 h exposure to the NF-κB inhibitor (BAY11-7082). Afterward, the supernatants were collected, and CCL2 levels were quantified at 24 h. ( D ) The expression of CXCL10, measured at 24 h. ( E ) The expression of IL-6, measured at 24 h. Statistical evaluations were conducted utilizing ANOVA (** p < 0.01).

Article Snippet: The TLR7 inhibitor (HY-124603, MCE, Princeton, NJ, USA), agonist (HY-117602, MCE, USA) and Myd88 inhibitor (HY-50937) were purchased from Med-Chem-Express.

Techniques: Translocation Assay, Western Blot, Incubation, Infection, Expressing