notch inhibitor dapt  (Tocris)

Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a) αSMA mRNA levels in HCAEC treated with TGFβ1 (10 ng/ml) or exposed to hypoxic conditions (5% O 2 ), in the absence or presence of moderate dose EtOH (25 mM, green bars) or high dose EtOH (100 mM, red bars) +/- the γ-secretase inhibitor DAPT (20 μM). (b) αSMA and fibronectin (FN) mRNA levels in HCAEC treated with TGFβ1 +/- the Notch ligand DLL4 (3 μg/ml) in the absence or presence of DAPT (20 μM). Data are mean±SEM, n=3. *p<0.05 vs control (no treatment), #p<0.05 vs TGFβ1 or hypoxia.

Article Snippet: Where indicated, cells were treated with Notch inhibitor DAPT (20 μM, Cat. #2634, Tocris,) or Notch ligand DLL4 (2 μg/ml, Cat. # 1506-D4, R&D Systems).

Techniques: Control

Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: HCAEC were treated with inflammatory cytokines (IL1β + TGFβ1) in the absence or presence of moderate dose ethanol (25 mM EtOH) or high dose ethanol (100 mM EtOH), with or without the gamma secretase inhibitor DAPT (20 μM), before Cdh5 expression was determined by immunocytochemistry. (a) Representative immunofluorescence images from 2 separate experiments, and (b) cumulative quantitative data of Cdh5 fluorescent staining. Data are mean±SEM, n=6. *p<0.05 vs control (no treatment, grp 1), #p<0.05 vs IL1β + TGFβ1 (grp 2).

Article Snippet: Where indicated, cells were treated with Notch inhibitor DAPT (20 μM, Cat. #2634, Tocris,) or Notch ligand DLL4 (2 μg/ml, Cat. # 1506-D4, R&D Systems).

Techniques: Expressing, Immunocytochemistry, Immunofluorescence, Staining, Control

Journal: Cell Reports Methods

Article Title: Rapid expansion of primary human vocal fold epithelial cells via targeted pathway inhibition and anchorage-independent sphere culture

doi: 10.1016/j.crmeth.2026.101310

Figure Lengend Snippet: Simultaneous TGF-β, ROCK, and Notch pathway inhibition promotes VFE proliferation while maintaining core epithelial phenotype (A) Effect of candidate small-molecule inhibitors on VFE proliferation. Cells (5 × 10 4 ) were incubated with 1 μM A-83-01 (TGF-β inhibitor), 10 μM Y-27632 (ROCK inhibitor), and 5 μM DAPT (Notch inhibitor), as indicated. Counts were performed at day 9; data are plotted as mean ± SEM ( n = 6); p values were obtained using mixed-model ANOVA with planned pairwise comparisons shown. Additional dose-response data for DAPT are presented in . (B) Experimental conditions used for subsequent experiments. Cells in the 3i-VFE condition were incubated with a three-molecule cocktail of 1 μM A-83-01, 10 μM Y-27632, and 5 μM DAPT; cells in the VFE condition were incubated with DMSO vehicle. (C) Bright-field images of live cells at passage 3; all cells exhibited cuboidal morphology over serial passages. Scale bar, 20 μm. (D) Single-passage growth curves and population doubling times. Data are plotted as mean ± SEM ( n = 4); p values were obtained using mixed-model ANOVA. (E) Flow cytometry data showing KRT14 and KRT19 expression. Positive/negative gates (versus unstained controls) are shown in gray. Data are plotted as mean ± SEM ( n = 3); p values were obtained using a paired t test. (F) RT-qPCR data showing TP63 , PROM1 , KIT , CDH1 , and MUC1 transcription; TUBB is reported as an additional reference standard. Data are presented as fold change relative to VFE (mean ± SEM; n = 3); p values were obtained using a paired t test. (G) Transepithelial electrical resistance. Data are plotted as mean ± SEM ( n = 6); the p value was obtained using a paired t test. (H) Flow cytometry data showing CD90 and MUC1 (also known as CD227) expression; VFFs are included as a non-epithelial cell control. High/low gates are shown in black. Data are plotted as mean ± SEM ( n = 3); p values were obtained using paired and unpaired t tests. Note that the 3i-VFE data (light blue) are largely superimposed on the VFE data (dark blue) in the dot plot.

Article Snippet: VFE were seeded in 24-well plates (Corning) at a density of 5 × 10 4 cells per well, cultured in VFE-orientated medium until ∼30% confluent, serum starved for 12 h, then incubated with various combinations of 1 μM TGF-β inhibitor A-83-01, 10 μM ROCK inhibitor Y-27632, and 5 μM Notch inhibitor DAPT (Selleck Chemicals), each prepared from a 1,000× stock solution in DMSO (Thermo Fisher).

Techniques: Inhibition, Incubation, Flow Cytometry, Expressing, Quantitative RT-PCR, Control

Journal: Science Advances

Article Title: 12R-HETE acts as an endogenous ligand for Nur77 in the intestines and regulates NKp46 + ILC3 development

doi: 10.1126/sciadv.adz8405

Figure Lengend Snippet: ( A ) Percentages of Ki67 + cells among RORγt + ILC3s are shown on the left. Absolute numbers of Ki67 + cells among RORγt + ILC3s are shown on the right. ( B ) Percentages of Ki67 + cells among NKp46 + ILC3s in the intestinal lamina propria. ( C ) Percentages of Ki67 + cells among NKp46 − ILC3s. ( D ) Numbers in flow plots represent percentages of T-bet + NKp46 + cells in the ILC3 gate. ( E ) Percentage of T-bet + cells among NKp46 + ILC3s. ( F ) IFN-γ MFI among NKp46 + ILC3s. ( G ) Percentage of T-bet + cells among DN ILC3s. ( H ) IFN-γ MFI among NKp46 + ILC3s. ( I ) Experimental design for the treatment of 12R-HETE and DAPT. SI LPLs were isolated from the 2-week-old mice. ( J ) Percentages and numbers of NKp46 + ILC3s were analyzed by flow cytometry after Notch signaling inhibitor treatment. ( K ) NKp46 MFI among NKp46 + ILC3. Data are means ± SEM; n = 5 to 6. Each point represents a biological replicate. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; n.s., not significant.

Article Snippet: In mechanistic experiments, mice were intraperitoneally injected with Notch inhibitors DAPT [MedChemExpress (MCE)] or Impdh1 inhibitors MPA (MCE) after treatment with PBS or 12R-HETE, and animals were euthanized after 10 days.

Techniques: Isolation, Flow Cytometry