PI3K/Akt/mTOR · target hub

EPHA5

Ephrin type-A receptor 5 (EPHA5)

EPHA5 is a Upstream RTK–Ras–MAPK node on the PI3K/Akt/mTOR axis. Receptor tyrosine kinase which binds promiscuously GPI-anchored ephrin-A family ligands residing on adjacent cells, leading to contact-dependent bidirectional signaling into neighboring cells. The signaling pathway downstream of the receptor is referred to as forward signaling wh… It is indexed under "Major receptor tyrosine kinases (selected)" on our pathway page for antibodies, inhibitors, and assay guidance.

Back to pathway overviewSearch: EPHA5UniProt (P54756)

Research notes

  • Combine with other genes in the same hub block via genetics or pharmacology to test non-redundant roles of EPHA5.
  • Annotate PIK3CA/PTEN and upstream RTK–Ras context when reading p-AKT, mTORC1/2, or lipid outputs.
  • Verify antibody clone, phospho-site, and stimulation; include KD/KO or inhibitor controls.

On the pathway hub (sections)

Activating lesions & amplification (quick reference)

Type / exampleDomain / context (brief)
Kinase domainKinase-domain activating lesions (RTK-dependent)
Fusion / amplificationFusion or amplification (cohort-dependent)
CN gainCopy gain and overexpression

Naming and prevalence vary by cohort and assay—annotate clinically with COSMIC, ClinVar, OncoKB, and datasheets; research context only.

Assay readouts for EPHA5

  • Receptor phosphorylation: p-RTK or p-Tyr blots—ligand time courses.
  • Downstream: p-AKT and p-ERK—partition PI3K vs MAPK dependence.
  • Genetics: KRAS/BRAF/PTEN context; bypass on resistance.
  • Inhibitor matrix: RTK/MEK/PI3K-selective agents.

EPHA5 experimental notes

Examples below reflect common literature and public resources (e.g., CCLE, DepMap)—validate genotypes, expression, and passage in your own stocks before committing assays.

[1] Cell lines commonly used for EPHA5 studies (examples)

  • Tool lines: A549, HepG2, PC-9, Ba/F3, HEK293T—screen by EPHA5 expression and pathway context (CCLE/DepMap).
  • Combine with neighbors (EGFR, ERBB2, ERBB3, ERBB4) via KD/pharmacology to test non-redundancy.
  • Overexpression/rescue in HEK293T supports mechanism and IP workflows.
  • Isogenic/CRISPR models separate pathway dependency from bypass survival.

[2] Cell samples for EPHA5 Western blot

  • Whole-cell lysates—optimize RIPA/NP-40 per antibody; phosphatase inhibitors for phospho blots.
  • Stimulation: serum starvation ± insulin/EGF or amino-acid withdrawal/refeed as relevant.
  • Controls: siRNA/shRNA, CRISPR KO, or inhibitors to validate band specificity.
  • Loading: BCA normalization; subcellular fractionation when needed.

[3] Tissue samples for EPHA5 Western blot

  • Matched tumor/adjacent frozen tissues after pathology review.
  • Mouse GEMM or xenografts—mind species antibody cross-reactivity.
  • Primary cells/PDCs when ethically approved.
  • FFPE needs specialized extraction; frozen tissue preferred for phospho work.

[4] Cell samples for EPHA5 immunoprecipitation

  • Tagged overexpression: HEK293T FLAG/HA-EPHA5 for complex capture.
  • Endogenous IP: high-expression lines; often ≥1–5×10⁶ cells per IP.
  • Stimulation: ligand or nutrient treatments enrich interactions—pilot time courses.
  • Controls: isotype IgG, empty vector, KD/KO negatives.

Human tissues and primary cells require ethics/IRB approval; tumors are heterogeneous—record histotype, site, and preservation conditions.

Bypass & related pathways

Models & genetics note

Overexpression vs endogenous EPHA5 can differ in dosage, splicing, and compartmentation—in organoids/PDX, record passage, matrix, and drug history.

Quick search presets

EPHA5EPHA5 antibodyPI3K inhibitormTOR inhibitor

EPHA5-related antibodies (keyword-biased)

Adds EPHA5 keyword bias atop the PI3K/Akt/mTOR antibody pool—if sparse, use presets above or global search.

Neighboring targets (same hub)

EGFRERBB2ERBB3ERBB4PDGFRAPDGFRBKITFLT1

External databases & resources

FAQ

This content supports research reagents and pathway education—not medical advice. Annotate mutations, drug indications, and protocols with authoritative databases, datasheets, and institutional oversight.

Last reviewed: 2026-05-18

Related on this site

See the PI3K/Akt/mTOR hub for neighboring nodes and assay guidance—cross-check MAPK, RTK, metabolism, and autophagy pages as needed.

PI3K/Akt/mTOR·MAPK/ERK·Receptor tyrosine kinase·Metabolism