Journal: Scientific Reports

Article Title: Pseudomonas aeruginosa -derived DnaJ functions as a novel immunomodulator inducing IFNβ via CME–SGK1–IRF3 axis in macrophages

doi: 10.1038/s41598-025-31281-x

Figure Lengend Snippet: DnaJ-induced IFNβ expression is regulated by TLR4. ( A , B , D , F – H , K , L ) dTHP-1 cells were pretreated for 1 h with: ( A , B ) IRAK4 IH (compound 26), ( D ) TLR2/4 IH (OxPAPC), ( F – H ) TLR4 IH (CLI-095; 3 µM in H ), or ( K , L ) MyD88 IH (T6167923). ( C , E , I , J ) Cells were transfected with: ( C ) siTLR10, ( E ) siTLR2, ( I ) siCD14, or ( J ) siMD2 for 48 h. Knockdown efficiency is shown in the right panel of ( C , E , I , J ). Following pretreatment or transfection, cells were treated with DnaJ (1 µg/ml) for 4 h ( A – G , I – L ) or for the indicated durations ( H ). IFNβ mRNA was quantified by qPCR; protein levels were analyzed by immunoblotting. The samples used for immunoblotting were obtained from the same experiment, and the original blots/gels are shown in the Supplementary Figure. Data in ( A – G , I – L ) are expressed as means ± SD ( n = 3). Immunoblots are representative of three experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001 vs. DnaJ treatment alone. Inhibitor (IH).

Article Snippet: OxPAPC (TLR2/4 inhibitor) and CLI-095 (TLR4 inhibitor) were purchased from InvivoGen (San Diego, CA, USA).

Techniques: Expressing, Transfection, Knockdown, Western Blot

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression.

doi: 10.3390/cells11223579

Figure Lengend Snippet: Figure 5. TLR4/NF-κB signaling cascade and ROS abundance are responsible for S100A8-induced NLRP3 inflammasome-dependent pyroptotic death in macrophages. (A) Western blot analysis of p65, p-p65, IKKα, and p-IKKα expression in THP-1 macrophages treated with GST-rhS100A8 or GST for 0, 30, 60 or 120 min. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the 0 min group (right panel). (B–E) THP-1 macrophages

Article Snippet: To investigate the endogenous PRR of S100A8, THP-1 differentiated macrophages were pretreated with the TLR4 inhibitor TAK-242 (10 μM, S7455, Selleck, Houston, Texas, USA) or the receptor for advanced end products (RAGE) inhibitor FPS-ZM1 (10 μM, S8185, Selleck, Houston, Texas, USA) for 1 h and then stimulated with rhS100A8 (5 μg/mL).

Techniques: Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Asprosin Exerts Pro-Inflammatory Effects in THP-1 Macrophages Mediated via the Toll-like Receptor 4 (TLR4) Pathway

doi: 10.3390/ijms24010227

Figure Lengend Snippet: Asprosin did not significantly affect Toll-like receptor 4 (TLR4) surface and intracellular expression. THP-1 macrophages were treated with either 10 nM asprosin, 100 nM asprosin or 100 ng/mL lipopolysaccharide (LPS). Cell surface expression of TLR4 was measured by flow cytometry after 4 and 24 h ( A ). Flow cytometry histograms display fluorescent intensity of the TLR4-PE stain ( x -axis) and cell count ( y -axis). Data were compared using one-way ANOVA and Dunnett’s multiple comparisons test (compared to control) ( n = 3). THP-1-cells (1 × 10 6 cells/well) were seeded and differentiated into macrophages in 6-well plates. Cells were then treated with either 100 nM asprosin or 100 ng/mL LPS for 4 and 24 h; collected and lysed for intracellular protein expression was measured by Western blotting ( B ). The presented Western data are representative of three independent experiments ( n = 3). CL: control; ASP: asprosin; LPS: lipopolysaccharide.

Article Snippet: To investigate the role of TLR4 in asprosin-induced inflammation, THP-1 derived macrophages were treated with 100 nM asprosin or 100 ng/mL LPS with and without a 1 h pre-treatment with 1 μM TAK-242 (R-Ethyl 6-(N-(2-chloro-4-fluorophenyl)sulfamoyl)cyclohex-1-enecarboxylate), a TLR4 signalling inhibitor (#6587; Tocris Bioscience, Bristol, UK).

Techniques: Expressing, Flow Cytometry, Staining, Cell Counting, Control, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Asprosin Exerts Pro-Inflammatory Effects in THP-1 Macrophages Mediated via the Toll-like Receptor 4 (TLR4) Pathway

doi: 10.3390/ijms24010227

Figure Lengend Snippet: Asprosin-induced inflammation in THP-1 macrophages is inhibited by TAK-242, a Toll-like receptor 4 (TLR4) inhibitor. ( A ) Tumour necrosis factor α (TNFα); ( B ) Interleukin-1β (IL-1β); and ( C ) IL-8 mRNA levels were measured by qRT PCR in THP-1 macrophages treated with 1 μM TAK-242 for 1 h, prior to 4 h stimulation with 100 nM asprosin or 100 ng/mL lipopolysaccharide (LPS). Data are presented as fold-change (mean ± SEM) in transcript. The experiments were repeated in three independent cultures. One-way ANOVA; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Secreted levels of TNFα ( D ), IL-1β ( E ), MCP-1 ( F ) and IL-8 ( G ) were measured by a human cytokine BioPlex array in cell supernatant of THP-1 macrophages treated with 1μM TAK-242 for 1 h, prior to 4 h stimulation with 100 nM asprosin or 100 ng/mL LPS. The experiments were repeated in three independent cultures. One-way ANOVA; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: To investigate the role of TLR4 in asprosin-induced inflammation, THP-1 derived macrophages were treated with 100 nM asprosin or 100 ng/mL LPS with and without a 1 h pre-treatment with 1 μM TAK-242 (R-Ethyl 6-(N-(2-chloro-4-fluorophenyl)sulfamoyl)cyclohex-1-enecarboxylate), a TLR4 signalling inhibitor (#6587; Tocris Bioscience, Bristol, UK).

Techniques: Quantitative RT-PCR

Journal: eLife

Article Title: Tumor-derived extracellular vesicles regulate tumor-infiltrating regulatory T cells via the inhibitory immunoreceptor CD300a

doi: 10.7554/eLife.61999

Figure Lengend Snippet: ( A ) Representative microscopy images of wild-type (WT) and Cd300a −/− bone marrow-derived dendritic cells (BMDCs) treated with pHrodo-labeled extracellular vesicles (EVs) to assess the localization of EVs (red) and early endosome antigen (EEA)-1 (green). Scale bar, 10 μm. Data are representative of two independent experiments. ( B ) Uptake of PKH-labeled tumor-derived EVs (TEVs) in WT ( n = 5) and Cd300a −/− BMDCs ( n = 5). ( C ) Representative microscopy images of WT and Cd300a −/− BMDCs treated with pHrodo-labeled exosomes to assess the localization of exosomes (green), TLR3 (red), and CD300a (blue). Scale bar, 10 μm. Data are representative of two independent experiments. ( D ) Quantitative RT-PCR analysis of Ifnb in WT and Cd300a −/− BMDCs treated with B16-derived exosomes in the presence of dimethyl sulfoxide (DMSO) (WT, n = 9; Cd300a −/− , n = 10), 100 nM TLR4 inhibitor ( n = 7 in each group), and 50 μM TLR3 inhibitor ( n = 6 in each group). ( E ) Quantitative RT-PCR analysis of Ifnb in WT, Cd300a −/− , ticam-1 −/− , and ticam-1 −/− ;Cd300a −/− mice-derived BMDCs treated with B16-derived EVs ( n = 5 in all group). ( F ) Representative immunoassay of WT and Cd300a −/− BMDCs left unstimulated (0 min) or stimulated for the indicated times with B16-derived exosomes, followed by immunoblot analysis of phosphorylated (p-) interferon regulatory factor 3 (IRF3) or total IRF3. Data are representative of two independent experiments. ( G and H ) Comparison of tumor growth and survival curves of B16 melanoma cells between ticam-1 −/− ( n = 6) and ticam-1 −/− ;Cd300a −/− ice ( n = 9) after inoculation of B16 melanoma. ( I and J ) Comparison of tumor growth and survival curves of B16 melanoma between MyD8 −/− ( n = 9) and MyD88 −/− ;Cd300a −/− mice ( n = 10) after inoculation of B16 melanoma. Data are given as means ± standard error of the means (SEMs). N.S.: not significant. *p < 0.05, **p < 0.01, and ***p < 0.001. p values were obtained by using the Student’s t -test ( B ), a two-way analysis of variance (ANOVA) followed by Bonferroni’s post-test ( D, E, G, and I ), and the log-rank test ( H and J ). Data were pooled from two ( B, E, and H ) or three ( D, I, and J ) independent experiments. Figure 5—source data 1. Source data for .

Article Snippet: Chemical compound, drug , TLR4 inhibitor (TAK-242) , Merck , Cat. #: 614,316 , .

Techniques: Microscopy, Derivative Assay, Labeling, Quantitative RT-PCR, Western Blot

Journal: eLife

Article Title: Tumor-derived extracellular vesicles regulate tumor-infiltrating regulatory T cells via the inhibitory immunoreceptor CD300a

doi: 10.7554/eLife.61999

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , TLR4 inhibitor (TAK-242) , Merck , Cat. #: 614,316 , .

Techniques: Purification, Sequencing, Recombinant, Isolation, Software

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: Resatorvid 3D molecular structures: (ChemDraw TM Ver. 16.0; Cambridge Soft, Cambridge, MA, USA).

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: Summary of partition coefficient (Log-P) for resatorvid at various pH (n = 3, mean ± standard deviation).

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: HPLC chromatogram of resatorvid. Format like 1e+5 means 1 ×10 5 .

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro Franz cell/Strat-M ® permeation profile of resatorvid: ( A ) 5% creams and lotion formulations; ( B ) 2.5% creams and lotion formulations; and ( C ) 1.25% creams and lotion formulations.

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro Franz cell/Strat-M ® permeation profile of resatorvid: ( A ) 5% simple solutions; ( B ) 2.5% simple solutions; and ( C ) 1.25% simple solutions.

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro Franz cell/Strat-M ® permeation profile of resatorvid: ( A ) 2.5% gel formulations and ( B ) 1.25% gel formulations.

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro drug membrane parameters of resatorvid topical creams and lotion formulations through Strat-M ® transdermal diffusion membrane ( n = 3, mean ± standard deviation). Lowercase letters indicate differences between formulations with the same concentration. Capital letters indicate difference between concentrations in the same formulation ( p < 0.05).

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro, Membrane, Diffusion-based Assay, Standard Deviation, Concentration Assay, Formulation, Cream

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro drug membrane parameters of resatorvid topical simple solutions through Strat-M ® transdermal diffusion membrane ( n = 3, mean ± standard deviation). Lowercase letters indicate differences between formulations with the same concentration. Capital letters indicate differences between concentrations in the same formulation ( p < 0.05).

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro, Membrane, Diffusion-based Assay, Standard Deviation, Concentration Assay, Formulation

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro cell viability of raw resatorvid on ( A ) human transformed keratinocytes (HaCaT) and ( B ) primary normal human epidermal keratinocytes (NHEKs).

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro, Transformation Assay

Journal: Pharmaceutics

Article Title: Design, Physicochemical Characterization, and In Vitro Permeation of Innovative Resatorvid Topical Formulations for Targeted Skin Drug Delivery

doi: 10.3390/pharmaceutics14040700

Figure Lengend Snippet: In vitro EpiDerm TM permeation profile of resatorvid: ( A ) 1.25% cream formulations; ( B ) 1.25% hydrogels formulations; and ( C ) 1.25% solutions.

Article Snippet: Resatorvid (TAK-242, molecular weight = 361.82 g/mol, C 15 H 17 ClFNO 4 S, >98% purity), shown in , was purchased from APAC Pharmaceutical LLC (Columbia, MD, USA).

Techniques: In Vitro, Cream

Journal: Translational Oncology

Article Title: IL-6 knockdown in a model of the human bone marrow, abrogates DNA damage induction in bystander cells post-chemotherapy induced cytokine release syndrome

doi: 10.1016/j.tranon.2024.102030

Figure Lengend Snippet: HS-5 viability and IL-6 expression following exposure to different dosages of resatorvid over 5 days. HS-5 cells were seeded and treated with range of resatorvid doses over 5 days (120 h). (A) HS-5 live cell counts were taken in every 24 h and (B) IL-6 secretion from HS-5 cells post-exposure to doses of resatorvid. (C) IL-6 expression from HS-5 cells exposed to 3 µM resatorvid with and without chemotherapy. Culture media were collected every 24 h following drug treatment and IL-6 level was analysed using ELISA. Data shows the mean ± SD ( n = 3) and significant differences shown as * p ≤ 0.05 and ** p ≤ 0.01 as determined by two-way ANOVA, Tukey's multiple comparisons test.

Article Snippet: Resatorvid (TAK-242; Stratech, UK) functions as a chemical inhibitor of IL-6 and TNFα.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Translational Oncology

Article Title: IL-6 knockdown in a model of the human bone marrow, abrogates DNA damage induction in bystander cells post-chemotherapy induced cytokine release syndrome

doi: 10.1016/j.tranon.2024.102030

Figure Lengend Snippet: Resatorvid treated HS-5 co-cultured with bystander TK6 cells. HS-5 cells were treated with resatorvid and chemotherapy, then co-cultured with TK6 bystander cells, separated by a culture insert. Conditioned media were collected on the subsequent day. (A) IL-6 expression from resatorvid treated HS-5 in isolation (grey bars) and from TK6 co-culture (white bars). (B) MN induction and RPD evaluation in bystander TK6 cells after co-culture. Data shows the mean ± SD ( n = 3) and significant differences are shown as * p ≤ 0.05, ** p ≤ 0.01 and **** p ≤ 0.0001 as determined by two-way ANOVA, Šídák's multiple comparisons test.

Article Snippet: Resatorvid (TAK-242; Stratech, UK) functions as a chemical inhibitor of IL-6 and TNFα.

Techniques: Cell Culture, Expressing, Isolation, Co-Culture Assay

Journal: Journal of Controlled Release

Article Title: Recent nanoengineered diagnostic and therapeutic advancements in management of Sepsis

doi: 10.1016/j.jconrel.2022.10.029

Figure Lengend Snippet: List of nanoformulations marketed and in clinical trials for management of infection and sepsis.

Article Snippet: TAK-242® , Takeda Global Research & Development Center, Inc. , Resatorvid emulsion , Sepsis , Phase IIa , [ ] .

Techniques: Clinical Proteomics, Infection, Emulsion, Polymer, Dispersion