tlr5  (Shanghai Korain Biotech Co Ltd)

Journal: bioRxiv

Article Title: Liver sinusoidal endothelial cells integrate metabolic and immune signals for MAPK-dependent BMP6 regulation and hepcidin induction

doi: 10.64898/2026.05.07.723498

Figure Lengend Snippet: (A) Bmp6 mRNA expression in primary LSECs treated with 5 ng/mL LPS or 2.5 μM heme for 6 h in the presence or absence of hepatocyte-conditioned medium. (B) Bmp6 mRNA expression in primary LSECs treated with various TLR ligands: Pam3CSK4 (TLR1/2), PGN (TLR2), Poly I:C (TLR3), LPS (TLR4), FLA-ST (TLR5), FSL1 (TLR2/6), R848 (TLR7/8), and ODN (TLR9) or vehicle control (NT, non-treated) for 6 h. (C-D) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of LPS for 6 h or with 5 ng/mL LPS for 2, 4, and 6 h. (E) Bmp6 mRNA expression in LSECs treated with 2.5 μM heme or protoporphyrin IX (PPIX) for 6 h. (F-G) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of heme for 6 h or treated with 2.5 μM heme for 2, 4, or 6 h. (H) Bmp6 mRNA expression in primary LSECs pre-treated with TAK242 (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS or 2.5 μM heme treatment for 6 h. (I) Transcription factor activity analysis by RNA-seq of LSECs treated with heme with respect to untreated controls (contrast-wise), in presence of hepatocyte-conditioned medium. (J) Bmp6 mRNA expression in primary LSECs pre-treated with CHX (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS treatment for 6 h. Cell culture experiments, except those in panel A, were always conducted in the presence of hepatocyte-conditioned medium. Gene expression levels were assessed by RT-qPCR, normalized to the housekeeping gene Rpl19 , and expressed as fold change relative to vehicle-treated controls. The dashed line (ut) represents the mRNA expression of LSECs treated with the conditions shown, in the absence of LPS or heme. Data are obtained from three or four independent experiments and displayed as mean ± SD. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA or Student’s t-test. CM, conditioned medium; PPIX, protoporphyrin IX; CHX, cycloheximide.

Article Snippet: The TLR ligands Pam3CSK4 (TLR2:1) (#tlrl-pms), PGN-SA (TLR2) (#tlrl-pgns2), Poly I:C (TLR3) (#tlrl-picw), FLA-ST (TLR5) (#tlrl-stfla), FSL1 (TLR2:TLR6) (tlrl-fsl), R848 (TLR7:8) (#tlrl-r848-1), ODN (TLR9) (#tlrl-1826) and the MAPK inhibitor SP600125 (#tlrl-sp60) were purchased form Invivogen and diluted in PBS (TLR ligands) or DMSO (SP600125).

Techniques: Expressing, Control, Activity Assay, Analysis, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Extensive genomic diversity in Desulfovibrio species reveals species-specific functional traits associated with disease

doi: 10.64898/2026.03.24.713149

Figure Lengend Snippet: a & b . Volcano plots showing COGs differentially enriched in CRC ( a ) and Crohn’s disease (CD) ( b ). Each point represents a COG. X-axis indicates COG prevalence in CD (or CRC) patients minus prevalence in healthy individuals. Y-axis shows log-transformed adjusted p-values (Fisher’s exact test, Benjamini-Hochberg correction for multiple testing). Dashed horizontal lines indicate significance threshold (adjusted p-value <0.25). c. Electron microscopy image of D. desulfuricans (MS020) shows visible flagella. d. Flagellated D. desulfuricans (MS020) and D. legallii (MS029) activate TLR5, while non-flagellated Desulfovibrio longus sp. nov. (MS012) showed no TLR5 activation. HEK-Blue TLR5 reporter cells were treated with bacterial culture supernatant filtrates. e. Recombinant flagellin proteins confirm flagellin-specific TLR5 activation. All but one of the recombinantly expressed flagellins (Fla2DD_MS020) significantly activated TLR5 (HEK-Blue TLR5 reporter cell line, linear mixed-effects model with Sidak comparison, stars represent significance level: * p<0.05; **** p<0.0001. The concentrations for FlaBS and FlaST are 0.5ng/ml and the concentrations for all recombinant Desulfovirbio flagellins are 1000 ng/ml. Abbreviations of the flagellins on the x-axis are defined as follows: FlaBS, flagellin from Bacillus subtilis; FlaST: flagellin from Salmonella typhimurium ; Fla1DD_MS021, flagellin 1 from D. desulfuricans (isolate MS021); Fla1DD_MS020, flagellin 1 from D. desulfuricans (isolate MS020); Fla2DD_MS020, flagellin 2 from D. desulfuricans (isolate MS020); Fla1DF: flagellin 1 from D. fairfieldensis (isolate MS026); Fla2DF: flagellin 2 from D. fairfieldensis (isolate MS026); Fla1DL: flagellin 1 from D. legallii (isolate MS029). f . Fla2DD_MS020 flagellin induces distinct gene expression patterns in murine small intestinal organoids. PCA analysis based on bulk RNAseq of flagellin-treated organoid experiments. WT = wild type organoids, Tlr5KO = TLR5 knock-out organoids. Unstimulated organoids were used as negative controls. g. Differentially expressed genes induced by Fla2DD_MS020 include upregulation of TGF-beta signaling and downregulation of genes involved in oxygen response (Wald test implemented in R package DESeq2, adjusted p <0.05).

Article Snippet: In parallel, HEK-Blue TLR5 reporter cell line (InvivoGen) reporting for murine TLR5 receptors and control cell line HEK-Blue Null1 without TLR gene overexpression (passages 7 to 9) were cultivated at 37 °C under 5% CO2 in Dulbecco MEM high glucose supplemented with 10% (v/v) fetal bovine serum, 10% (v/v) antibiotic antimycotic solution, 1mM L-Glutamine and specific antibiotics according to manufacturers’ recommendations.

Techniques: Transformation Assay, Electron Microscopy, Activation Assay, Recombinant, Comparison, Gene Expression, RNA sequencing, Knock-Out

Journal: Neoplasia (New York, N.Y.)

Article Title: Microbe-associated molecular patterns differentially mediate carcinogenic alterations of the breast tissue in the context of obesity

doi: 10.1016/j.neo.2026.101284

Figure Lengend Snippet: LPS and flagellin induce DNA double-strand breaks in breast acini in a TLR-dependent manner. A. Representative comet images from neutral comet assays performed on 3D cultures of breast acini treated with vehicle (control), 1 μg/ml LPS, 1 μg/ml LTA, and 10 ng/ml flagellin for 72 h. Bleomycin (BLM; 20 mU/ml; 2h) was used as positive control. B. Comet assay quantification. n=300 acini from 3 biological replicates. C. Representative images for immunofluorescence staining of 53BP1 (red) in acini treated with MAMPs ± TLR inhibitors. Nuclei are counterstained with DAPI (blue). D-F. Quantification of 53BP1 foci numbers in acini treated with LPS ± TLR4 inhibitor (D), with LTA ± TLR2 inhibitor (E) or with flagellin ± TLR5 inhibitor (F). ns is not significant. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Antibodies against TLR4 (Santa Cruz; sc-293072), TLR5 (Santa Cruz; sc-518106), TLR2 (Santa Cruz; sc-21795), NF-κB p65 (Cell Signaling Technology; 8242), LEDGF (Cell Signaling Technology; 2088), Lamin B (Abcam; Ab16048), and α-tubulin (Cell Signaling Technology; 2144) were used.

Techniques: Control, Positive Control, Single Cell Gel Electrophoresis, Immunofluorescence, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Microbe-associated molecular patterns differentially mediate carcinogenic alterations of the breast tissue in the context of obesity

doi: 10.1016/j.neo.2026.101284

Figure Lengend Snippet: Blockade of TLR4 and TLR5 mitigates DNA damage in the mammary glands. A. Schematic of the experiment. B-D-F. Representative images for TLR2 (B), TLR4 (D), and TLR5 (F) IHC signals in mammary glands of mice treated with saline, mismatch morpholino (MM) and the corresponding TLR morpholino (TLR M). Hematoxylin was used as a counterstain. Image analysis and quantification of TLR2/4/5-DAB positive areas is shown in the graphs (C,E,G). H. Representative comet images of the neutral comet assay performed on mammary cells treated with vehicle, MM, and TLR morpholinos. I. Quantification of comet assay results for the aforementioned mice groups. ns is not significant. *P<0.05, **P<0.01.

Article Snippet: Antibodies against TLR4 (Santa Cruz; sc-293072), TLR5 (Santa Cruz; sc-518106), TLR2 (Santa Cruz; sc-21795), NF-κB p65 (Cell Signaling Technology; 8242), LEDGF (Cell Signaling Technology; 2088), Lamin B (Abcam; Ab16048), and α-tubulin (Cell Signaling Technology; 2144) were used.

Techniques: Saline, Neutral Comet Assay, Single Cell Gel Electrophoresis

Journal: bioRxiv

Article Title: Transcriptome-based lead generation, ligand- and structure-based prioritization and experimental validation of TLR5-activating molecules

doi: 10.64898/2026.02.25.707690

Figure Lengend Snippet:

Article Snippet: TLR5 secreted in the culture supernatants were measured using Human Toll-like Receptor 5, TLR5 ELISA kit (BT LAB, #E0374Hu) according to the manufacturer’s protocol.

Techniques: Control

Journal: bioRxiv

Article Title: Transcriptome-based lead generation, ligand- and structure-based prioritization and experimental validation of TLR5-activating molecules

doi: 10.64898/2026.02.25.707690

Figure Lengend Snippet: This figure shows all the H-bond interactions between robust CMap-generated leads and TLR5 which fall below 3 Å. The minimum 3 H-bond interactions between CMap-generated leads and TLR5 were showed by both Glide module and HADDOCK tool in each complex.

Article Snippet: TLR5 secreted in the culture supernatants were measured using Human Toll-like Receptor 5, TLR5 ELISA kit (BT LAB, #E0374Hu) according to the manufacturer’s protocol.

Techniques: Generated

Journal: bioRxiv

Article Title: Transcriptome-based lead generation, ligand- and structure-based prioritization and experimental validation of TLR5-activating molecules

doi: 10.64898/2026.02.25.707690

Figure Lengend Snippet: (A) CAL 27 cells were incubated with the CMap-generated leads: Cytarabine (200µM, 100µM, 50µM), Azacytidine (200µM, 100µM, 50µM), Penicillin (200µM, 100µM, 50µM), Streptozotocin (400µM, 200µM, 100µM), Piceatannol (400µM, 200µM, 100µM) and flagellin (5ng/ml) for 24h. (B) CAL 27 cells were incubated with the CMap-generated leads: Fenoterol (400µM, 200µM, 100µM), ABT-751 (200µM, 100µM, 50µM), Ganciclovir (200µM, 100µM, 50µM), Kinetin-riboside (100µM, 50µM, 25µM), and flagellin (5ng/ml) for 24h. Bar graph representing the relative expression levels of TLR5 with respect to the untreated control. Fold change is calculated by the signal of the treated sample divided by the signal of the control. If the fold change is >1, TLR5 expression is upregulated whereas fold change <1 is taken as downregulated. All values are expressed as mean ± SEM.

Article Snippet: TLR5 secreted in the culture supernatants were measured using Human Toll-like Receptor 5, TLR5 ELISA kit (BT LAB, #E0374Hu) according to the manufacturer’s protocol.

Techniques: Incubation, Generated, Expressing, Control

Journal: bioRxiv

Article Title: Transcriptome-based lead generation, ligand- and structure-based prioritization and experimental validation of TLR5-activating molecules

doi: 10.64898/2026.02.25.707690

Figure Lengend Snippet:

Article Snippet: TLR5 secreted in the culture supernatants were measured using Human Toll-like Receptor 5, TLR5 ELISA kit (BT LAB, #E0374Hu) according to the manufacturer’s protocol.

Techniques: Control

Journal: bioRxiv

Article Title: Transcriptome-based lead generation, ligand- and structure-based prioritization and experimental validation of TLR5-activating molecules

doi: 10.64898/2026.02.25.707690

Figure Lengend Snippet: This figure shows all the H-bond interactions between robust CMap-generated leads and TLR5 which fall below 3 Å. The minimum 3 H-bond interactions between CMap-generated leads and TLR5 were showed by both Glide module and HADDOCK tool in each complex.

Article Snippet: TLR5 secreted in the culture supernatants were measured using Human Toll-like Receptor 5, TLR5 ELISA kit (BT LAB, #E0374Hu) according to the manufacturer’s protocol.

Techniques: Generated

Journal: bioRxiv

Article Title: Transcriptome-based lead generation, ligand- and structure-based prioritization and experimental validation of TLR5-activating molecules

doi: 10.64898/2026.02.25.707690

Figure Lengend Snippet: (A) CAL 27 cells were incubated with the CMap-generated leads: Cytarabine (200µM, 100µM, 50µM), Azacytidine (200µM, 100µM, 50µM), Penicillin (200µM, 100µM, 50µM), Streptozotocin (400µM, 200µM, 100µM), Piceatannol (400µM, 200µM, 100µM) and flagellin (5ng/ml) for 24h. (B) CAL 27 cells were incubated with the CMap-generated leads: Fenoterol (400µM, 200µM, 100µM), ABT-751 (200µM, 100µM, 50µM), Ganciclovir (200µM, 100µM, 50µM), Kinetin-riboside (100µM, 50µM, 25µM), and flagellin (5ng/ml) for 24h. Bar graph representing the relative expression levels of TLR5 with respect to the untreated control. Fold change is calculated by the signal of the treated sample divided by the signal of the control. If the fold change is >1, TLR5 expression is upregulated whereas fold change <1 is taken as downregulated. All values are expressed as mean ± SEM.

Article Snippet: TLR5 secreted in the culture supernatants were measured using Human Toll-like Receptor 5, TLR5 ELISA kit (BT LAB, #E0374Hu) according to the manufacturer’s protocol.

Techniques: Incubation, Generated, Expressing, Control