tlr3 inhibitor cu cpt 4a  (MedChemExpress)

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Differentially expressed genes (DEGs) in full-thickness bladder tissues from HIC patients and controls identified via RNA sequencing (|log₂FC|≥2; adjusted p < 0.05, two-sided Wald test using DESeq2). Gene Ontology (GO) ( B ) and KEGG ( C ) enrichment analyses of DEGs (two-sided hypergeometric test with FDR correction). D UMAP visualization of single-cell transcriptomic landscape of urothelium, with urothelial cells (UCs) identified by KRT19 and UPK1A expression. GO ( E ) and KEGG ( F ) analyses of upregulated DEGs in UCs (two-sided hypergeometric test with FDR correction). GO ( G ) and KEGG ( H ) analyses of downregulated DEGs in UCs (two-sided hypergeometric test with FDR correction). I , J Gene Set Enrichment Analysis (GSEA) of altered pathways in UCs (two-sided permutation test with 1000 permutations, with multiple testing corrected using the FDR). The exact p value for TLR signaling of UCs in panel I is 0.0009. K Expression of Toll-like receptor (TLR) subtypes in UCs from HIC bladders. mRNA ( L ) and protein ( M ) expression levels of TLR3 in isolated urothelium from control and HIC patients ( n = 7 control vs. 10 HIC; data show median (IQR); two-sided Mann–Whitney U test; ns not significant; Bar for panel M: 1 cm). N Immunostaining showing the distribution of TLR3 protein in the urothelium of patients with HIC ( n = 7 control vs. 10 HIC; one section and field per patient; Bar: 100 μm). IHC immunohistochemistry, HL Hunner lesions, U urothelium, LP lamina propria (blue line), M muscularis. Source data are provided as a Source Data file.

Article Snippet: TLR3 inhibitor group: Same intravesical TCDCA instillation as above, followed by intraperitoneal injection of a TLR3 inhibitor (CU-CPT 4a, MCE, Cat. HY-108473) at 1 mg/kg , twice per week for 2 weeks.

Techniques: RNA Sequencing, Expressing, Isolation, Control, MANN-WHITNEY, Immunostaining, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Experimental workflow of E. avium colonization in antibiotic-pretreated mice (Abx-mice, n = 6 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . B Increased abundance of E. avium confirmed by metagenomic sequencing ( n = 6 per group). C Evaluation of voiding behavior following E. avium transplantation ( n = 6 per group; one measurement per mouse). D Measurement of mechanical pain threshold post-transplantation ( n = 6 per group). E Representative histological analysis of bladder tissues after E. avium transplantation ( n = 6 per group; one section and field per mouse). F , G Quantification of blood and urinary bile acids using comprehensive targeted bile acid profiling ( n = 6 per group). H Cell viability of human urothelial cells (HUCs) following TCDCA or TUDCA treatment ( n = 3 independent experiments). I Expression of ZO-1, TNF-α, and TLR3 following TCDCA (200 µM) exposure ( n = 3 independent experiments). J Experimental workflow of intravesical instillation of TCDCA in rats ( n = 5 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . K Evaluation of voiding function after TCDCA instillation ( n = 5 per group; each rate measured once). L Assessment of mechanical pain threshold following TCDCA exposure ( n = 5 per group). M Representative histology of bladder tissues post-instillation ( n = 5 per group; one section per rat, one field quantified per section). For panels ( B – H , K , L ): data are presented as median (IQR). Statistical analysis was performed using the two-sided Mann–Whitney U test. IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: TLR3 inhibitor group: Same intravesical TCDCA instillation as above, followed by intraperitoneal injection of a TLR3 inhibitor (CU-CPT 4a, MCE, Cat. HY-108473) at 1 mg/kg , twice per week for 2 weeks.

Techniques: Sequencing, Transplantation Assay, Expressing, MANN-WHITNEY, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A , B Expression of tight junction protein ZO-1 and inflammatory marker TNF-α after TLR3 intervention in TCDCA-pretreated human urothelial cells (HUCs) ( n = 3 independent experiments). C Transepithelial resistance (TER) changes following TLR3 intervention in TCDCA-pretreated HUCs ( n = 3 independent experiments). D , E Expression of ZO-1 and TNF-α after pentosan polysulfate sodium (PPS) administration in TCDCA-pretreated HUCs ( n = 3 independent experiments). F TER changes following PPS intervention in TCDCA-pretreated HUCs ( n = 3 independent experiments). G Experimental workflow showing TLR3 inhibitor and PPS administration to rats following intravesical TCDCA instillation ( n = 5 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . H Assessment of voiding function (one measurement per rat) and pain threshold after TLR3 inhibition ( n = 5 per group). I Representative histological images of bladder tissues post-TLR3 inhibition ( n = 5 per group; one section and field per rat). For panels ( B , C , E , F , H ): data are presented as median (IQR). Statistical analysis was performed using the Kruskal–Wallis test. ns not significant, IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: TLR3 inhibitor group: Same intravesical TCDCA instillation as above, followed by intraperitoneal injection of a TLR3 inhibitor (CU-CPT 4a, MCE, Cat. HY-108473) at 1 mg/kg , twice per week for 2 weeks.

Techniques: Expressing, Marker, Inhibition, Immunohistochemistry

Journal: Journal of Extracellular Vesicles

Article Title: Cancer‐Specific RNA Modifications in Tumour‐Derived Extracellular Vesicles Promote Tumour Growth

doi: 10.1002/jev2.70083

Figure Lengend Snippet: m6A levels in EVs regulate inflammatory responses via TLR8 in macrophages. Differentiated THP‐1 cells (dTHP‐1) were pretreated with or without CU‐CPT‐9a and CU‐CPT‐4a. (A) Whole‐cell lysates obtained from dTHP‐1 cells were subjected to Western blot analysis using an anti‐phospho p65 antibody, anti‐p65 antibody and anti‐β‐actin antibody. Representative images from three independent experiments are shown. Conditioned medium from dTHP‐1 cells treated with CCD‐841‐CoN or HT29 small EVs was used for ELISA. TNF‐α (B, left and C, left), and IL‐6 (B, right and C, right). Values are presented as the mean ± SEM for each group. One‐way ANOVA post hoc Tukey's test; * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant. dTHP‐1 cells were pre‐transfected with or without TLR8 siRNA. (D) Whole‐cell lysates obtained from dTHP‐1 cells treated with CCD‐841‐CoN small EVs or HT29 small EVs were subjected to Western blot analysis using an anti‐phospho p65 antibody, anti‐p65 antibody and anti‐β‐actin antibody. Representative images from three independent experiments are shown. Conditioned medium was used for the ELISA. TNF‐α (E, left) and IL‐6 (E, right). Values are presented as the mean ± SEM for each group. One‐way ANOVA post hoc Tukey's test; ** p < 0.01, *** p < 0.001, **** p < 0.0001. dTHP‐1 cells were pre‐transfected with or without TLR3 siRNA. (F) Whole‐cell lysates obtained from dTHP‐1 cells treated with CCD‐841‐CoN small EVs or HT29 small EVs were subjected to Western blot analysis using an anti‐phospho p65 antibody, anti‐p65 antibody and anti‐β‐actin antibody. Representative images from three independent experiments are shown. Conditioned medium was used for ELISA. TNF‐α (G, left) and IL‐6 (G, right). Values are presented as the mean ± SEM for each group. One‐way ANOVA post hoc Tukey's test; *** p < 0.001, **** p < 0.0001; ns, not significant. (H) HT29 cells were cultured in a conditioned medium from dTHP‐1 cells transfected with or without TLR8 siRNA and CCD‐841‐CoN small EVs or HT29 small EVs. Values are presented as the mean ± SEM for each group. One‐way ANOVA post hoc Tukey's test; * p < 0.05, ** p < 0.01. HT29‐derived EVs‐RNA were pretreated with or without rALKBH5 and introduced into dTHP‐1 cells transfected with or without TLR8 siRNA. TNF‐α (I) and IL‐6 (J) concentrations were measured via ELISA using conditioned medium from dTHP‐1 cells. Values are presented as the mean ± SEM for each group. One‐way ANOVA post hoc Tukey's test; * p < 0.05, *** p < 0.001, **** p < 0.0001, ns, not significant. See also Figure .

Article Snippet: TLR8 agonist (Motolimod), TLR3 agonist (Poly(I:C)), TLR8 inhibitor (CU‐CPT9a) and TLR3 inhibitor (CU‐CPT‐4a) were purchased from MedChemExpress (Monmonth Junction, NJ, USA). dTHP‐1 cells (1 × 10 5 cells) were seeded in 24‐well plates and treated with inhibitors for 48 h simultaneously with EVs or agonists.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Transfection, Cell Culture, Derivative Assay