Notch Signaling Pathway Diagram Maker
Generate Notch signaling pathway diagrams between two cells, from ligand binding and the ADAM10 and γ-secretase cuts to NICD and CSL (RBPJ) at HES and HEY genes, plus lateral inhibition and T-ALL. Made for cell biology.
Notch signaling pathway examples
Real renders of four Notch signaling prompts: the fully labeled pathway, a five-label overview, lateral inhibition in the inner ear and a blank worksheet. Click one to load its prompt and edit it.
Two cells in contact, a receptor cut twice
Read down from the sending cell to the nucleus below.
- 1
DLL1 → Notch receptor
The ligand on the sending cell binds Notch on the receiving cell.
- 2
Ligand endocytosis
The sending cell pulls the bound ligand in, tugging on Notch.
- 3
S2 and S3 cleavage
ADAM10 cuts outside the membrane; γ-secretase then cuts inside.
- 4
NICD, CSL (RBPJ), MAML
In the nucleus, NICD joins CSL and MAML to turn on HES1 and HEY1.
One cell keeps its Delta, the other loses it
Read left to right, from Cell A across into Cell B.
- 1
Delta on A → Notch on B
Cell A, high in Delta, activates Notch on its neighbour, Cell B.
- 2
NICD → HES1
NICD released in Cell B switches on HES1.
- 3
HES1 on B's Delta gene
HES1 turns down B's own Delta, so the difference between them grows.
Notch figures from receptor structure to disease
The figures above show one ligand–receptor pair and lateral inhibition in the inner ear, and the table below sets Notch against Wnt. These are the other Notch figures developmental biology, vascular biology and hematology papers draw.
Notch receptor domain structure
The receptor as a linear map, N-terminus outside. Extracellular: a long row of EGF-like repeats, 36 in NOTCH1, with the ligand-binding site at repeats 11–12; then the negative regulatory region — three Lin12–Notch repeats and the heterodimerization domain — which hides the S2 site until a ligand pulls. Intracellular: the RAM domain and seven ankyrin repeats, which bind CSL, nuclear localization signals and the C-terminal PEST domain. Mark the three cuts on it: S1 by furin, S2 by ADAM10, S3 by γ-secretase within the transmembrane domain.
Lateral induction and boundary formation
The opposite of the lateral inhibition in the second figure: Notch activation raises ligand, usually JAG1, in the receiving cell, so the signal spreads and a whole field of cells takes the same fate — the prosensory domain of the inner ear and the smooth muscle layers of arteries are the standard examples. At a boundary, Fringe makes Notch in dorsal cells respond to Delta and not to Serrate, so signaling is confined to a line, as at the dorsal–ventral border of the Drosophila wing disc.
Tip and stalk cells in angiogenesis
A sprouting vessel with two endothelial cells. VEGF-A acting on VEGFR2 raises DLL4 in the leading tip cell; DLL4 activates NOTCH1 in the stalk cell behind it, which lowers VEGFR2 and raises VEGFR1, so that cell stays behind and divides to build the stalk. JAG1 in stalk cells competes with DLL4 and keeps the selection dynamic. Blocking DLL4 gives many tip cells and a dense tangle of vessels that carry little blood.
NOTCH1 in the thymus and in T-ALL
Two panels. In the thymus, DLL4 on the cortical epithelium activates NOTCH1 in incoming progenitors and commits them to the T-cell lineage instead of the B-cell lineage. In T-ALL the same receptor is stuck on: draw the mutations onto the receptor map — heterodimerization-domain mutations that let it be cleaved without ligand, PEST truncations and FBXW7 loss that keep NICD from being degraded. A rare translocation, t(7;9), puts a truncated NOTCH1 under the control of the T-cell receptor β locus.
Notch in the intestinal crypt
Notch decides between the two lineages of the gut lining. Paneth cells present DLL1 and DLL4 to the neighbouring Lgr5⁺ stem cells and progenitors; in Notch-on cells HES1 represses ATOH1 and they become absorptive enterocytes, while Notch-off cells express ATOH1 and become goblet, Paneth or enteroendocrine cells. Blocking γ-secretase turns the crypt over to goblet cells, which is the mechanism behind the diarrhea in γ-secretase inhibitor trials.
Inherited Notch disorders
Each syndrome drawn at its own node. Alagille syndrome: mostly JAG1, a few NOTCH2 mutations; paucity of bile ducts, heart defects such as peripheral pulmonary stenosis, butterfly vertebrae and posterior embryotoxon. CADASIL: NOTCH3 mutations that change the number of cysteines in an EGF-like repeat, in vascular smooth muscle; subcortical strokes, migraine with aura and dementia. Hajdu–Cheney syndrome: NOTCH2 truncations that remove the PEST domain. Spondylocostal dysostosis: DLL3 and other genes of the somite segmentation clock. NOTCH1 mutations also occur in bicuspid aortic valve and aortic valve calcification.
What is the Notch signaling pathway?
The Notch signaling pathway is a contact-dependent pathway: a Delta-like or Jagged ligand on one cell binds the Notch receptor on its neighbour, ADAM10 and γ-secretase cut the receptor, and the freed Notch intracellular domain (NICD) enters the nucleus to switch on HES and HEY genes with CSL (RBPJ) and MAML. The receptor fragment itself carries the signal, with no kinase cascade or second messenger in between.
Key facts
- Mammals have four Notch receptors and five canonical ligands. NOTCH1–4 are activated by the Delta-like ligands DLL1 and DLL4 and the Jagged ligands JAG1 and JAG2; DLL3 works mainly by cis-inhibition rather than by activating Notch.
- Notch is cut three times. Furin cleaves the new receptor at S1 in the Golgi, making the mature heterodimer; after ligand binding, ADAM10 cuts at S2 just outside the membrane, and γ-secretase, whose catalytic subunit is presenilin, cuts at S3 inside it to release NICD.
- The ligand has to pull. Binding alone is not enough: the ubiquitin ligases Mindbomb or Neuralized tag the bound ligand, the sending cell endocytoses it, and that pull unfolds the receptor's negative regulatory region so ADAM10 can reach the S2 site.
- CSL (RBPJ) is a switch. Without NICD it sits on target genes with co-repressors and histone deacetylases; NICD displaces them and recruits MAML and histone acetyltransferases such as p300 to turn HES1, HES5, HEY genes and MYC on.
- Each signal is short-lived. CDK8 phosphorylates NICD's PEST domain and the ubiquitin ligase FBXW7 marks it for destruction, resetting the pathway — PEST and FBXW7 mutations that stop this are common in T-cell acute lymphoblastic leukemia.
Components of the Notch signaling pathway and what each one does
The molecules a labeled Notch pathway diagram is normally expected to show, in the order the signal passes through them. This is the default label set unless you ask for a shorter one.
| Component | What it does | Where to draw it |
|---|---|---|
| DSL ligand (DLL1, DLL4, JAG1, JAG2) | Transmembrane ligand that binds Notch on the neighbouring cell | In the sending cell's membrane, facing the gap |
| Mindbomb or Neuralized | Ubiquitin ligases that tag the bound ligand for endocytosis, which pulls on the receptor | Inside the sending cell, at the ligand's tail |
| Notch receptor (NOTCH1–4) | Single-pass receptor, held as a two-part heterodimer after S1 cleavage by furin | Spanning the receiving cell's membrane |
| Fringe | Golgi glycosyltransferase that extends O-fucose on Notch's EGF repeats, favouring Delta over Jagged | In the receiving cell's Golgi (optional) |
| ADAM10 (S2 cleavage) | Metalloprotease that sheds the extracellular domain once the pull exposes the S2 site, leaving NEXT | Just outside the receiving cell's membrane |
| γ-Secretase (S3 cleavage) | Intramembrane protease — presenilin with nicastrin, PEN-2 and APH-1 — that cuts NEXT to release NICD | Within the membrane bilayer |
| NICD | Notch intracellular domain; carries the signal itself into the nucleus | Arrow from the membrane through the cytoplasm to the nucleus |
| CSL (RBPJ) with co-repressors | DNA-binding protein that keeps target genes off while NICD is absent | On the DNA in the nucleus |
| MAML (Mastermind-like) | Co-activator recruited by NICD and CSL; brings in p300 | Stacked on NICD and CSL on the DNA |
| HES and HEY genes | Main targets; they encode repressors that carry out Notch's effects | Downstream of the complex, at the bottom |
Courses differ in how many of these they expect. An introductory figure stops at ligand, receptor, NICD and target genes; a cell biology course adds ADAM10, γ-secretase, CSL (RBPJ) and MAML; advanced courses add furin, Fringe and Mindbomb. Sources also differ on the S2 enzyme: ADAM17 cuts Notch in the test tube, but ligand-induced cleavage in living cells depends on ADAM10.
Notch vs Wnt signaling: what is the difference?
Notch signaling needs direct contact — a membrane-bound ligand on one cell triggers cuts in the receptor on the next, and the receptor fragment itself goes to the nucleus — whereas canonical Wnt signaling uses a secreted ligand that saves β-catenin from its destruction complex. Both steer cell fate in development and stem cells, and both are rewired in cancer.
| Notch signaling | Wnt (canonical) signaling | |
|---|---|---|
| Ligand | Membrane-bound DSL ligands: DLL1, DLL4, JAG1, JAG2 | Secreted, lipid-modified WNT glycoproteins — 19 in humans |
| Range | Contact-dependent (juxtacrine): only touching cells | Short and long range, as a gradient around the source |
| Receptor | NOTCH1–4, single-pass transmembrane | One of 10 Frizzled receptors with the LRP5/6 co-receptor |
| How the signal passes | Proteolysis: S2 by ADAM10, S3 by γ-secretase | The destruction complex (APC, Axin, GSK3, CK1) is switched off |
| Messenger to the nucleus | NICD, a fragment of the receptor | β-catenin, a cytoplasmic protein spared from degradation |
| Nuclear partner | CSL (RBPJ) with MAML | TCF/LEF |
| Off state | CSL (RBPJ) represses targets with co-repressors | β-catenin is phosphorylated and destroyed by the proteasome |
| Signature cancer | NOTCH1 mutations in more than 50% of T-ALL | A Wnt-pathway mutation in about 90% of sporadic colorectal cancers |
Draw Notch as two cells touching, with the ligand fixed in one membrane; draw Wnt with the ligand free in the space between cells. The two pathways cross-talk — NICD can act on Wnt signaling — but keep their nuclear partners apart: CSL for Notch, TCF/LEF for Wnt.
Common questions about the Notch signaling pathway
What should a labeled Notch signaling pathway diagram include?
The standard set is the signal-sending cell with a DSL ligand (Delta-like or Jagged), the signal-receiving cell with the Notch receptor, S2 cleavage by ADAM10, S3 cleavage by γ-secretase, the released NICD, and in the nucleus CSL (RBPJ), MAML and the HES/HEY target genes. Many figures add S1 cleavage by furin in the Golgi and the co-repressors CSL holds before NICD arrives.
What are the steps of Notch signaling?
Five steps: a ligand on one cell binds Notch on the next; the sending cell endocytoses the ligand and pulls; ADAM10 cuts Notch at S2; γ-secretase cuts it at S3 inside the membrane; and NICD enters the nucleus, where it joins CSL (RBPJ) and MAML to switch on target genes. Before any of this, furin has already cut the new receptor once, at S1 in the Golgi.
What does γ-secretase do in Notch signaling?
γ-Secretase makes the last cut, S3, inside the membrane, releasing NICD from the stub that ADAM10 leaves behind. It is a four-part complex — presenilin 1 or 2 as the catalytic subunit, with nicastrin, PEN-2 and APH-1 — and it also cleaves amyloid precursor protein, the substrate linked to Alzheimer's disease.
What is lateral inhibition in Notch signaling?
Lateral inhibition is how Notch splits a group of equivalent cells into two fates: a cell that sends slightly more Delta activates Notch in its neighbours, Notch signaling lowers their own Delta, and the difference grows until the first cell takes one fate and the cells around it another. In the inner ear this leaves each hair cell surrounded by supporting cells.
What is cis-inhibition of Notch?
Cis-inhibition is Notch being blocked by ligand in its own cell: a Delta or Jagged ligand in the same membrane binds the receptor and stops it responding to ligand on neighbours. DLL3 seems to act only this way, and together with activation in trans it pushes a cell into a clear sending or receiving state.
What does Fringe do to Notch?
Fringe enzymes — Lunatic, Manic and Radical Fringe — extend the O-fucose sugars on Notch's EGF repeats in the Golgi, which makes NOTCH1 respond more to Delta-like ligands than to Jagged. The modification tunes the signal; it is not required for the receptor to mature.
How is Notch signaling involved in cancer?
Activating NOTCH1 mutations occur in more than 50% of T-cell acute lymphoblastic leukemias (T-ALL), mostly in the heterodimerization domain, which lets the receptor fire without ligand, and in the PEST domain, which keeps NICD from being degraded. In other tissues, such as squamous cell carcinomas, Notch acts as a tumor suppressor instead.
What are γ-secretase inhibitors used for?
γ-Secretase inhibitors (GSIs) block the S3 cut and so shut Notch signaling down; the FDA approved one, nirogacestat, for progressing desmoid tumors in November 2023. An early GSI trial in T-ALL was halted by severe diarrhea — the gut lining depends on Notch, and the enzyme has many substrates besides Notch.
Sources
- Signaling by NOTCH1 — pathway R-HSA-1980143 — Reactome
- Notch signaling pathway — KEGG PATHWAY hsa04330 — KEGG
- Notch signaling pathway: architecture, disease, and therapeutics (Zhou et al., 2022) — Signal Transduction and Targeted Therapy
- FDA approves nirogacestat for desmoid tumors (2023) — U.S. Food and Drug Administration
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