Journal: Nature

Article Title: Resolving the fibrotic niche of human liver cirrhosis at single cell level

doi: 10.1038/s41586-019-1631-3

Figure Lengend Snippet: a , Dotplot: ligand-receptor interactions between SAMΦ (n=10 human livers) and SAMes (n=7 human livers). X-axis, ligand (red) and cognate receptor (blue); y-axis, cell populations expressing ligand (red) and receptor (blue); circle size, P value (permutation test); colour (red, high; yellow, low), means of average ligand and receptor expression levels in interacting subpopulations. b , Representative immunofluorescence images, fibrotic niche. Left, TREM2 (red), PDGFB (white), PDGFRα (green), DAPI (blue), arrows TREM2 + PDGFB + cells. Right, TNFRSF12A (red), TNFSF12 (white), PDGFRα (green), DAPI (blue), arrows TNFRSF12A + PDGFRα + cells, scale bars 50μm. c to d , HSC proliferation assay: y-axis, area under curve (AUC) of % change in HSC number over time (hours), Mean±SEM, one-way ANOVA and Tukey. c , Control, TNFSF12, anti-TNFRSF12A, isotype control, all n=3. d , Vehicle, PDGF-BB, Crenolanib, all n=3. e , Dotplot: ligand-receptor interactions between SAEndo (n=7 human livers) and SAMes (n=7 human livers). X-axis, ligand (red) and cognate receptor (blue); y-axis, cell populations expressing ligand (red) and receptor (blue); circle size, P value (permutation test); colour (red, high; yellow, low), means of average ligand and receptor expression levels in interacting subpopulations. f , Representative immunofluorescence image, fibrotic niche. NOTCH3 (red), DLL4 (white), PDGFRα (green), DAPI (blue), arrows NOTCH3 + PDGFRα + cells, scale bar 50μm. g , Endothelial cell JAG1 flow cytometry: healthy (n=3) or cirrhotic (n=9) liver, representative histogram (left), mean fluorescence intensity (MFI, right), Mean±SEM, Mann-Whitney two-tailed. h , Cirrhotic endothelial cell and HSC co-culture, Notch inhibitor Dibenzazepine (DBZ). Representative immunofluorescence images (left), Collagen 1 (magenta), PECAM1 (green), DAPI (blue). Digital pixel analysis (right); collagen 1 area, n=3, Mean±SEM, RM one-way ANOVA and Tukey. n , HSC gene knockdown: control (n=7) or NOTCH3 (n=7) siRNA, qPCR of stated gene, expression relative to mean expression of control siRNA, Mean±SEM, Mann-Whitney two-tailed.

Article Snippet: After 2h, all growth factor supplements were removed and cells were cultured for a further 72 hours in endothelial basal media containing 10% heat-inactivated human serum ± Notch-signalling inhibitor Dibenzazepine (10μM, Bio-Techne, 4489/10) or vehicle (DMSO) control.

Techniques: Expressing, Immunofluorescence, Proliferation Assay, Flow Cytometry, Fluorescence, MANN-WHITNEY, Two Tailed Test, Co-Culture Assay

Journal: Cancer Science

Article Title: In vivo effects of chemotherapy on oncogenic pathways in colorectal cancer

doi: 10.1111/cas.14077

Figure Lengend Snippet: Impact of long‐term fluorouracil (5‐FU) chemotherapy and MAPK and NOTCH inhibition on colon cancer. A, Schema and experimental schedule for xenografting, treatment and tumor analysis. B, Long‐term treatment effects of 5‐FU, selumetinib (AZD) and dibenzazepine (DBZ), and their combination compared to vehicle (control) on SW480 and SW1222 colon cancer xenografts. Data are shown as growth curves and represent mean ± SE. n ≥ 3 for each treatment group. C, Representative H&E‐stained tissue sections of xenograft tumors after long‐term treatment, as indicated. Scale bars, 2 mm. D, Quantification of tumor necrosis in treated colon cancer xenografts. Error bars indicate mean ± SD. P values are t test results. n ≥ 3

Article Snippet: Irinotecan was obtained from Pfizer (Berlin, Germany), selumetinib (AZD) from Selleck Chemicals (Munich, Germany), and dibenzazepine (DBZ) from Axon Medchem (Groningen, the Netherlands).

Techniques: Inhibition, Control, Staining

Journal: Cancer Science

Article Title: In vivo effects of chemotherapy on oncogenic pathways in colorectal cancer

doi: 10.1111/cas.14077

Figure Lengend Snippet: Proliferation and apoptosis in colon cancer after long‐term therapy. A, Representative immunostaining for Ki‐67 and (B) quantification of Ki‐67‐positive tumor cells in colon cancer xenografts after indicated chemotherapeutic and/or inhibitor treatments in comparison to vehicle (control). C, Representative immunostaining for cleaved (Cl.) Caspase‐3 and (D) quantification of cleaved Caspase‐3‐positive tumor cells in colon cancer xenografts after indicated chemotherapeutic and/or inhibitor treatments, or vehicle (control) treatment. Error bars indicate mean ± SD. P ‐values are t test results. n ≥ 3. Scale bars, 50 μm. AZD, selumetinib; DBZ, dibenzazepine; 5‐FU, fluorouracil

Article Snippet: Irinotecan was obtained from Pfizer (Berlin, Germany), selumetinib (AZD) from Selleck Chemicals (Munich, Germany), and dibenzazepine (DBZ) from Axon Medchem (Groningen, the Netherlands).

Techniques: Immunostaining, Comparison, Control

Journal: Cancer Science

Article Title: In vivo effects of chemotherapy on oncogenic pathways in colorectal cancer

doi: 10.1111/cas.14077

Figure Lengend Snippet: Expression of thymidylate synthase on long‐term therapy in colon cancer xenografts. A, Representative immunostaining for thymidylate synthase and (B) quantification of thymidylate synthase positive tumor cells in colon cancer xenografts after indicated chemotherapeutic and/or inhibitor treatments, or vehicle (control) treatment. Error bars indicate mean ± SD. P ‐values are t test results. n ≥ 3. Scale bars, 50 μm. AZD, selumetinib; DBZ, dibenzazepine; 5‐FU, fluorouracil

Article Snippet: Irinotecan was obtained from Pfizer (Berlin, Germany), selumetinib (AZD) from Selleck Chemicals (Munich, Germany), and dibenzazepine (DBZ) from Axon Medchem (Groningen, the Netherlands).

Techniques: Expressing, Immunostaining, Control

Journal: Cell death & disease

Article Title: Enhancer remodeling activates NOTCH3 signaling to confer chemoresistance in advanced nasopharyngeal carcinoma.

doi: 10.1038/s41419-023-06028-z

Figure Lengend Snippet: Fig. 5 Chemical inhibitor targeting Notch signaling restores chemosensitivity of NPC cells in vitro. A Combination index (CI) of Dibenzazepine (DBZ) and paclitaxel (PTX) in 5–8 F R cells. The synergistic effect is demonstrated by CI value < 1.0. CI values were calculated with CalcuSyn software. Cells treated with indicated concentration of DBZ or PTX, single or in combination for 96 h. Cell viability was determined by CellTiter Glo reagent. B Colony formation assay of 5–8 F R cells treated with 25 μM DBZ in the presence or absence of 3.0 nM PTX for 8 days. All the following experiments used the same concentrations of DBZ or PTX, unless otherwise notified. PTX + DBZ (P + D). C Cell relative proliferation curve of 5–8 F R cells with different treatments. Data are shown as means ± SD (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001. D Tumorsphere formation assay of 5–8 F R cells treated with Notch signaling inhibitor in the presence or absence of PTX for 12 days. Scale bar, 400 μm. E Sub-G1 population analysis in 5–8 F R cells treated with Notch signaling inhibitor, PTX or both for 72 h. Data are shown as means ± SD (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001. F Annexin-V-PI staining to assess the apoptotic ratios of 5–8 F R cells under single or combined treatment for 72 h. Left, representative images of FACS analysis; Right, percentages of apoptotic cells in treatment groups as observed with flow cytometry. Bars represent the means ± SD, n = 3. *P < 0.05; **P < 0.01; ***P < 0.001. G Immunoblotting analysis of NICD3 under different treatments. Cells were collected after 72 h treatment. Results shown are representative image from three experiments.

Article Snippet: Dibenzazepine (DBZ), SGC-CBP30 and dCBP1 were purchased from TargetMol.

Techniques: In Vitro, Software, Concentration Assay, Colony Assay, Tube Formation Assay, Staining, Cytometry, Western Blot