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, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR

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: 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: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: a , Quantitative PCR of TLR5 from whole liver biopsy tissue, grouped by histological stage (ctrl n=5, steatosis n=9, MASH F1-2 n=5, MASH F3-4 n=5). b , Representative images of immunofluorescence of human liver biopsy tissue (n=12) showing TLR5 (green), albumin (grey), α-SMA (red), Hoescht (blue) in control (top row) and MASH with F3 fibrosis (bottom row) (Scale bar 200µm). c , NanoString CosMX analysis of explant liver tissue from MASH advanced fibrosis. Red dots represent TLR5 expressing cells. d , Zoomed representative images of immunofluorescence staining of human liver biopsy tissue showing TLR5 (green) and α-SMA (red) in MASH F3 (n=12,scale bar 100µm). e , f , Kaplan–Meier curves for liver-related events and all-cause mortality stratified by hepatic TLR5 expression in the SteatoSITE cohort. Error bars represent mean±standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Real-time Polymerase Chain Reaction, Immunofluorescence, Control, Expressing, Staining, Standard Deviation

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: a , Publicly-available single nucleus RNA sequencing data from 47 liver biopsies from individuals with different stages of MASLD (Gribben et al. Nature 2024, PMID 38778114 ) were analysed. Pie chart demonstrates proportion of TLR5 expressing single cells (n=1609) that co-expressed albumin (ALB, n=1495), equating to 92.9% derived from albumin-expressing hepatocytes. b , Liver TLR5 mRNA expression in SteatoSITE cohort divided according to histological MASLD disease severity. c , Liver TLR4 mRNA expression in SteatoSITE cohort divided according to histological MASLD disease severity. d, Liver TLR2 mRNA expression in SteatoSITE cohort divided according to histological MASLD disease severity. e , All-cause mortality by TLR5 expression in SteatoSITE cohort with MASH fibrosis stage 0 – 2. f , All-cause mortality by TLR5 expression in SteatoSITE cohort with MASH fibrosis stage 3 – 4. Kaplan–Meier curves for all-cause mortality stratified by hepatic TLR5 expression. P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: RNA Sequencing, Expressing, Derivative Assay

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet:

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Expressing

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: Primary hepatocytes treated with oleic acid and palmitic acid (OA PA), flagellin or combination (OA PA + flagellin) for 24 hours a, in the presence or absence of 1µM TH1020 (TLR5 inhibitor) , 5µg/mL Pab hTLR4 (TLR4 inhibitor), 5µg/mL Pab hTLR2 (TLR2 inhibitor), b - f in the presence or absence of 1µM TH1020. a , IL-8 concentration (n=3–4 replicates per condition). b , Representative images of lipid deposition quantified using BODIPY 493/503 staining (green), nuclei stained with DAPI (blue) via confocal microscopy (Scale bars 50μm) and BODIPY quantified according to relative intensity/DAPI. Experiments performed in triplicates for each condition. c - e Expression of fatty acid oxidation enzymes: c, very long chain specific acyl-CoA dehydrogenase – ACADVL, d, medium-chain specific acyl-CoA dehydrogenase - ACADM and e, long-chain 3-hydroxyl-CoA dehydrogenase – HADHA. f , DNA damage-inducible transcript 3 (DDIT3) concentration (n=2 independent experiments). Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple. ANOVA, analysis of variance; Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Concentration Assay, Staining, Confocal Microscopy, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: a , HepG2 cells, b , Huh7 cells, treated with oleic acid and palmitic acid (OA PA), flagellin or combination (OA PA + flagellin) for 24 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor). Representative images of lipid deposition quantified using BODIPY 493/503 staining (green), nuclei stained with DAPI (blue) via confocal microscopy (Scale bars 50µm). Experiments performed in triplicates for each condition. c – h, Expression of fatty acid oxidation enzymes ( c – e in HepG2 cells, f – h in Huh7 cells) of very long chain specific acyl-CoA dehydrogenase – ACADVL, medium-chain specific acyl-CoA dehydrogenase - ACADM and long-chain 3-hydroxyl-CoA dehydrogenase – HADHA following treatment with OA PA, flagellin or combination (OA PA flagellin) for 24 hours in the presence or absence of 1µM TH1020 (n=2 independent experiments). Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Staining, Confocal Microscopy, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: DNA damage-inducible transcript 3 (DDIT3) concentration in a , HepG2 cells; b , Huh7 cells, treated with oleic acid and palmitic acid (OA PA), flagellin or combination (OA PA + flagellin) for 24 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) (n=2 independent experiments). c , d , IL-8 concentration in c , HepG2 cells, d , Huh7 cells, treated with OA PA for 24 hours in the presence or absence of 27µM CU-CPT4a (TLR3 inhibitor), 5µg/mL PAb hTLR4 (TLR5 inhibitor), 1µM TH1020 (n=3 independent experiments). e , Flagellin concentration conditioned media from primary hepatocytes treated for 24 hours treatment with OA PA compared to control (n=3 independent experiments). f , IL-8 concentration in primary hepatocytes treated with 250µM oleic acid 125µM palmitic acid (OA PA), 25ng/mL flagellin or combination (OA PA + flagellin) for 24 hours (n=2 independent experiments). g , IL-8 concentration in HEK cells treated with OA PA, flagellin, and combination (OA PA + flagellin) for 24 hours (n=3 independent experiments). Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Concentration Assay, Control, Standard Deviation

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: a , HEK-Blue™ hTLR5 cells treated with oleic acid and palmitic acid (OA PA), flagellin for 24 hours. TLR5 ligand activity assessed by optical density measurements of SEAP production. n=3 independent experiments. b , Primary hepatocytes treated with OA PA for 24 hours in the presence or absence of fatty acid transporter protein inhibitor cocktail (10µM obeticholic acid (FATP5 inhibitor), 80µM SML2148 (CD36 inhibitor), 50µM Lipofermata (FATP2 inhibitor)). Representative images of lipid deposition quantified using BODIPY 493/503 staining (green), nuclei stained with DAPI (blue) via confocal microscopy (Scale bars 50µm). Experiments performed in triplicate for each condition. c – e , IL-8 concentrations in c, Huh7 cells treated with OA PA for 24 hours in the presence or absence of 16µM SML2148, 10µM Lipofermata, 2µM obeticholic acid, combination of 16µM SML2148, 10µM Lipofermata and 2µM obeticholic acid in a cocktail, 1µM TH1020 (TLR5 inhibitor) with 10µM Lipofermata (n=3 independent experiments). d , Primary hepatocytes treated with OA PA for 24 hours in the presence or absence of 40µM Pepinh-TRIF (TRIF inhibitor), 40µM TJ-M2010-5 (MyD88 inhibitor) (n=2 independent experiments) and e , HepG2 cells treated with OA PA for 24 hours in the presence or absence of 5µM (5Z)-7-Oxozeaenol (TAK1 inhibitor) (n=3 independent experiments). f, Schematic diagram depicting TLR5 signalling in hepatocytes (created using Biorender). Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Activity Assay, Staining, Confocal Microscopy, Standard Deviation

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: a , b , IL-8 concentration in a , HepG2 cells; b , Huh7 cells, treated with oleic acid and palmitic acid (OA PA) for 24 hours in the presence or absence of 40µM Pepinh-TRIF (TRIF inhibitor) (n=3 independent experiments). c , IL-8 concentration in HepG2 cells treated with OA PA for 24 hours in the presence or absence of 40µM TJ-M2010-5 (MyD88 inhibitor) (n=3 independent experiments). IL-8 concentration in Huh7 cells treated with OA PA for 24 hours in the presence or absence of 5µM (5Z)-7-Oxozeaenol (TAK1 inhibitor) (n=3 independent experiments). e , TGFβ1 concentration in LX2 cells following 24 hours treatment with conditioned media from primary hepatocytes treated with OA PA, flagellin in the presence or absence of 1µM TH1020 (TLR5 inhibitor) (n=3 replicates per condition). P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: Normalised RNA counts for TLR5 and TLR4 extracted from primary human hepatic stellate cells isolated from non-fibrotic, non-steatotic human livers (n=19). b , TGFβ1 concentration c , Pro-collagen 1⍺1 concentration, in LX2 cells treated with oleic acid and palmitic acid (OA PA), flagellin for 24 hours (n=3 independent experiments). d , TGFβ1 concentration, e , Pro-collagen 1⍺1 concentration, in LX2 cells treated with conditioned media from HepG2 cells treated with OA PA, flagellin in the presence or absence of 1µM TH1020 (TLR5 inhibitor) (n=3 independent experiments). Schematic diagram created using Biorender. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Isolation, Concentration Assay, Standard Deviation

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and Collagen III quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: For TLR5 inhibition, liver slices were incubated with 1 μM TLR5 antagonist, TH1020, (HY-116961-5, MedChemexpress) for 16 hours.

Techniques: Cell Culture, Immunohistochemistry, Standard Deviation