Journal: Journal of Translational Medicine

Article Title: Esophageal microbial dysbiosis impairs mucosal barrier integrity via toll-like receptor 2 pathway in patients with gastroesophageal reflux symptoms

doi: 10.1186/s12967-024-05878-1

Figure Lengend Snippet: The potential mechanisms that esophageal microbial dysbiosis induced GER symptoms. Esophageal microbial dysbiosis induced esophageal barrier dysfunction via LPS-TLR2-IL-6 pathway, thus triggering GER symptoms. Abbreviation: G-: Gram-negative; LPS: lipopolysaccharide; TLR: toll-like receptor; IL: interleukin; GER: gastroesophageal reflux

Article Snippet: Various concentrations of human IL-6 protein (MCE, HY-P7044; 0 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml and 400 ng/ml) was used to investigate its direct effects on claudin-1 expression and intercellular spaces (Supplementary Fig. B).

Techniques: Reflux

Journal: Journal of Translational Medicine

Article Title: Esophageal microbial dysbiosis impairs mucosal barrier integrity via toll-like receptor 2 pathway in patients with gastroesophageal reflux symptoms

doi: 10.1186/s12967-024-05878-1

Figure Lengend Snippet: Effects of LPS stimulation on HEECs. A . The mRNA and protein expression of TLR2 following LPS exposure (200 µg/ml). TLR2 mRNA expression: Ctrl vs. LPS = 1.0 ± 0.1 vs. 1.9 ± 0.1, P < 0.05; TLR2 protein expression: Ctrl vs. LPS = 0.7 ± 0.4 vs. 1.5 ± 0.1, P < 0.01. B . The mRNA and protein expression of claudin-1 following LPS exposure (200 µg/ml). Claudin-1 mRNA expression: Ctrl vs. LPS = 1.0 ± 0.0 vs. 0.4 ± 0.0, P < 0.001; claudin-1 protein expression: Ctrl vs. LPS = 1.5 ± 0.1 vs. 1.0 ± 0.1, P < 0.05. C . The intercellular space shown by H&E staining following LPS exposure (200 µg/ml): Ctrl vs. LPS = 0.4 ± 0.1 μm vs. 1.5 ± 0.2 μm, P < 0.001. D . The mRNA and protein expression of IL-6 following LPS exposure (200 µg/ml). IL-6 mRNA expression: Ctrl vs. LPS = 1.0 ± 0.2 vs. 3.9 ± 0.0, P < 0.001; IL-6 protein expression: Ctrl vs. LPS = 189.3 ± 5.4 pg/ml vs. 473.5 ± 7.9 pg/ml, P < 0.001. Abbreviation: Ctrl: control; LPS: lipopolysaccharide; IL: interleukin; DIS: dilated intercellular space; HEEC: human esophageal epithelial cell line; *: P < 0.05; **: P < 0.01; ***: P < 0.001

Article Snippet: Various concentrations of human IL-6 protein (MCE, HY-P7044; 0 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml and 400 ng/ml) was used to investigate its direct effects on claudin-1 expression and intercellular spaces (Supplementary Fig. B).

Techniques: Expressing, Staining, Control

Journal: Journal of Translational Medicine

Article Title: Esophageal microbial dysbiosis impairs mucosal barrier integrity via toll-like receptor 2 pathway in patients with gastroesophageal reflux symptoms

doi: 10.1186/s12967-024-05878-1

Figure Lengend Snippet: TLR2 blockade attenuated the effects of LPS. A . The mRNA and protein expression of IL-6 after blocking TLR2 (5 µg/ml). IL-6 mRNA expression: Ctrl vs. LPS vs. LPS + IgA vs. IgA = 1.0 ± 0.3 vs. 9.0 ± 1.3 vs. 4.8 ± 0.4 vs. 1.0 ± 0.1, P < 0.001; IL-6 protein expression: Ctrl vs. LPS vs. LPS + IgA vs. IgA = 62.4 ± 11.8 pg/ml vs. 210.6 ± 14.7 pg/ml vs. 61.1 ± 30.5 pg/ml vs. 49.9 ± 12.4 pg/ml, P < 0.001. B . The mRNA and protein expression of claudin-1 after blocking TLR2 (5 µg/ml). Claudin-1 mRNA expression: Ctrl vs. LPS vs. LPS + IgA vs. IgA = 1.0 ± 0.0 vs. 0.8 ± 0.0 vs. 0.9 ± 0.0 vs. 0.9 ± 0.0, P < 0.001; claudin-1 protein expression: Ctrl vs. LPS vs. LPS + IgA vs. IgA = 1.0 ± 0.1 vs. 0.6 ± 0.1 vs. 1.0 ± 0.1 vs. 0.9 ± 0.1, P < 0.001. C . The intercellular space shown by H&E staining after blocking TLR2 (5 µg/ml). Ctrl vs. LPS vs. LPS + IgA vs. IgA = 0.4 ± 0.2 μm vs. 1.7 ± 0.5 μm vs. 0.5 ± 0.2 μm vs. 0.6 ± 0.2 μm, P < 0.001. Abbreviation: Ctrl: control; LPS: lipopolysaccharide; TLR: toll-like receptor; ***: P < 0.001

Article Snippet: Various concentrations of human IL-6 protein (MCE, HY-P7044; 0 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml and 400 ng/ml) was used to investigate its direct effects on claudin-1 expression and intercellular spaces (Supplementary Fig. B).

Techniques: Expressing, Blocking Assay, Staining, Control

Journal: Journal of Translational Medicine

Article Title: Esophageal microbial dysbiosis impairs mucosal barrier integrity via toll-like receptor 2 pathway in patients with gastroesophageal reflux symptoms

doi: 10.1186/s12967-024-05878-1

Figure Lengend Snippet: IL-6 production is crucial for LPS-induced effects. A . The mRNA and protein expression of claudin-1 following IL-6 stimulation (400 ng/ml). Claudin-1 mRNA expression: Ctrl vs. IL-6: 1.0 ± 0.0 vs. 0.8 ± 0.0, P < 0.001; claudin-1 protein expression: Ctrl vs. IL-6: 1.2 ± 0.3 vs. 0.8 ± 0.1, P < 0.01. B . The intercellular space shown by H&E staining following IL-6 stimulation (400 ng/ml). Ctrl vs. IL-6: 0.4 ± 0.3 μm vs. 1.3 ± 0.3 μm, P < 0.01. C . The mRNA and protein expression of claudin-1 after treated with IL-6 antibody (0.1 ng/ml). Claudin-1 mRNA expression: Ctrl vs. LPS vs. LPS ± IL-6 IgG vs. IgG = 1.0 ± 0.0 vs. 0.8 ± 0.0 vs. 1.1 ± 0.0 vs. 1.0 ± 0.0, P < 0.001; claudin-1 protein expression: Ctrl vs. LPS vs. LPS ± IL-6 IgG vs. IgG = 1.2 ± 0.1 vs. 0.8 ± 0.1 vs. 1.2 ± 0.3 vs. 1.1 ± 0.1, P < 0.05. D The intercellular space shown by H&E staining after treated with IL-6 antibody (0.1 ng/ml). Ctrl vs. LPS vs. LPS ± IL-6 IgG vs. IgG = 0.2 ± 0.1 μm vs. 1.4 ± 0.0 μm vs. 0.4 ± 0.1 μm vs. 0.2 ± 0.1 μm, P < 0.001. Abbreviation: Ctrl: control; IL: interleukin; LPS: lipopolysaccharide; DIS: dilated intercellular space; *: P < 0.05; **: P < 0.01; ***: P < 0.001

Article Snippet: Various concentrations of human IL-6 protein (MCE, HY-P7044; 0 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml and 400 ng/ml) was used to investigate its direct effects on claudin-1 expression and intercellular spaces (Supplementary Fig. B).

Techniques: Expressing, Staining, Control

Journal: bioRxiv

Article Title: TGF-β1-induced differentiation enhances chemotherapy response in metastatic colorectal cancer organoids

doi: 10.64898/2026.06.04.730067

Figure Lengend Snippet: Cytokine modulation of differentiation. A scRNAseq analysis at 11 dps showing that CRC0322, CRC0327, and CRC1502 cells exhibit IL6R expression. B ELISA measuring the concentration of IL-6 sR, IL-6 and TGF-β1 in the conditioned media of CRC organoids over time. Curves show the average of 2 independent experiments. C-E Flow cytometry analysis of CRC0322 (C), CRC0327 (D), and CRC1502 (E) of organoids after 3 days of incubation with either IL-6 (50 ng/ml) or TGF-β1 (5 ng/ml). Histograms show the mean + SD of the percentage of GABRA2 positive cells for 4-7 independent experiments. Statistical significances were analyzed by one-sample t-tests of the log 2 fold change relative to the Control, with p-values corrected by Bonferroni adjustment. dps: days-post-seeding. *** p <0.001.

Article Snippet: For cytokine treatments, organoids were cultured for 11 days, after which human IL-6 (50 ng/mL; #130-093-931, Miltenyi Biotec) or human TGF-beta 1 Recombinant Protein (5 ng/mL; #100-21-10UG, PeproTech®) was added for 3 days prior to collection.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Flow Cytometry, Incubation, Control

Journal: bioRxiv

Article Title: TGF-β1-induced differentiation enhances chemotherapy response in metastatic colorectal cancer organoids

doi: 10.64898/2026.06.04.730067

Figure Lengend Snippet: Effect of IL-6 and TGF-β1 on CRC organoids and response to chemotherapy. A smFISH of LGR5, KRT20 and Ki67 in CRC0322 after 3 days with IL-6 (50 ng/ml) or TGF-β1 (5 ng/ml). Scale bars = 20 μm. B Histogram shows the percentage of positive LGR5, KRT20 and Ki67 cells. C Scatter plots show percentage of positive cells per organoid (each dot represents an organoid). The median of (LGR5+;KRT20+) cells in organoids were (41.7;37.5)% CONTROL, (38.7;40.5)% IL-6, (10.7;72.3)% TGF-β1 (CONTROL vs IL-6 p -value = 0.7279, CONTROL vs TGF-β1 p -value = 0.0001, PERMANOVA test). D, F Volcano plots of bulk RNA-seq of CRC0322 after 3 days with IL-6 (D) or TGF-β1 (F). E, G Scatter plots show upregulated and downregulated genes (p<0.05, |log 2 FC|>1; color scale) for IL-6 (E) or TGF-β1 (G) treated organoids and log 2 FC in the scRNAseq 11 dps sample within cells classified as Stem or Differentiated versus all other cells. Genes belonging to either module are circled. H Scheme of the organoid-initiation assay. I Histogram shows the mean + SD of number of live cells relative to control (CTL) after IL-6 or TGF-β1 treatment of CRC0322 from 3 independent experiments pooled together (ANOVA followed by Dunett’s test versus CTL). J Images of organoids after replating 1000 live cells from previously treated organoids (scale bar = 2000 μm). Histogram shows the mean + SD of the number of organoids relative to the CTL from 3 independent experiments pooled together (ANOVA followed by Dunett’s test versus CTL). K, L Representative MTS viability assay of CRC0322 (K) and CRC 1502 (L) after 7 days with oxaliplatin plus IL-6 or TGF-β1 (replicates=6) (Brown-Forsythe ANOVA followed by Dunett’s T3 test (CRC0322); ANOVA followed by Dunett’s test (CRC1502) of oxaliplatin treated conditions versus oxalipatin CTL). Values were normalized to the untreated CTL. Experiments were repeated 3 times, obtaining similar results. Fluorescent images: LGR5 (cyan), Ki67 (magenta), KRT20 (yellow), and DAPI (gray). dps: days-post-seeding; ns: not significant; FC: Fold change. **p<0.01; ****p<0.0001.

Article Snippet: For cytokine treatments, organoids were cultured for 11 days, after which human IL-6 (50 ng/mL; #130-093-931, Miltenyi Biotec) or human TGF-beta 1 Recombinant Protein (5 ng/mL; #100-21-10UG, PeproTech®) was added for 3 days prior to collection.

Techniques: Control, RNA Sequencing, Viability Assay