PI3K/Akt/mTOR · target hub

RPS6

40S ribosomal protein S6 (RPS6)

RPS6 is a mTORC1 translational output on the PI3K/Akt/mTOR axis. RPS6 participates in PI3K/Akt/mTOR signaling—see UniProt and primary literature for isoforms and tissue context. It is indexed under "S6K & ribosomal protein S6 axis" on our pathway page for antibodies, inhibitors, and assay guidance.

Back to pathway overviewSearch: RPS6UniProt (A2A3R6)

Research notes

  • Combine with other genes in the same hub block via genetics or pharmacology to test non-redundant roles of RPS6.
  • 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)

Genetic lesions & expression context (quick reference)

Type / exampleDomain / context (brief)
Expression contextTissue/cell-type–dependent expression
Rare variantsSporadic variants—require functional validation
Pathway couplingCo-occurs with neighboring nodes on the hub

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

Assay readouts for RPS6

  • Translation: p-S6K, p-RPS6, 4E-BP phospho/supershift, eIF4F assembly.
  • Polysome profiling or puromycin labeling (advanced readouts).
  • mTORC1 block: rapamycin or ATP-competitive mTOR inhibitors—time courses.
  • Cross-read MAPK/MNK axes where substrates overlap.

RPS6 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 RPS6 studies (examples)

  • Tool lines: HEK293T, HeLa, MCF-7, HCT116, A549—screen by RPS6 expression and pathway context (CCLE/DepMap).
  • Combine with neighbors (RPS6KB1, RPS6KB2) 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 RPS6 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 RPS6 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 RPS6 immunoprecipitation

  • Tagged overexpression: HEK293T FLAG/HA-RPS6 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 RPS6 can differ in dosage, splicing, and compartmentation—in organoids/PDX, record passage, matrix, and drug history.

Quick search presets

RPS6RPS6 antibodyPI3K inhibitormTOR inhibitor

RPS6-related antibodies (keyword-biased)

Adds RPS6 keyword bias atop the PI3K/Akt/mTOR antibody pool—if sparse, use presets above or global search.

Neighboring targets (same hub)

RPS6KB1RPS6KB2

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