Rho Activator I

Rho Activator I

Product Uses Include

  • Activator for Rho pathway
  • Study the effects of Rho activation on cell motility
  • Study the effects of Rho activation on the rearrangement of the actin cytoskeleton.
  • Investigate the effects of Rho activation with respect to cross talk to other signal transduction pathways

    Rho Activator I(Cat. # CN01) is useful for efficient activation of RhoA, RhoB, and RhoC in a variety of cultured cells.  The reagent activates Rho proteins in fibroblasts, neurons, epithelial, endothelial, and hematopoietic cells as well as other primary and immortalized lines.  Cells treated with the activator can be subjected to any one of a number of assays that indicate an increase in Rho activity, including focal adhesion or stress fiber staining (Cat. # BK005) and Rho activity assays by G-LISA™ (Cat. # BK124).  See Figure 1 for example of Rho activation measured by the G-LISA assay. 

    There are many activators of RhoA,B and C proteins in mammalian cells. Commonly used ones are calf serum (1), lysophosphatidic acid (LPA)(1) and calpeptin (2). Through years of experience in Rho activation assay, Cytoskeleton Inc. has identified calpeptin as a compound that activates many cell types and has a long timespan of activation for ease of use so this is the active component in Rho Activator I Cat.# CN01. Note: Calpeptin is used as an inhibitor of calpain, but it also inhibits myosin light chain phosphorylation which is connected to stress fiber formation and hence possibly to RhoA activation. 



    1. Ren, XD., Kiosses, WB., & Schwartz, MA.  Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton. EMBO J. 18: 578-585 (1999).
    2. Schoenwaelder, SM., & Burridge, K.  Evidence for a calpeptin-sensitive protein-tyrosine phosphatase upstream of the small GTPase Rho.  J. Biol. Chem. 274: 14359-14367 (1999)

    For product Datasheets and MSDSs please click on the PDF links below.   For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

    AuthorTitleJournalYearArticle Link
    Faisal, Syed M et al.Spatiotemporal Insights into Glioma Oncostream Dynamics: Unraveling Formation, Stability, and Disassembly PathwaysAdvanced Science2024ISSN 2198--3844
    Moztarzadeh, Sina et al.Cortactin is in a complex with VE-cadherin and is required for endothelial adherens junction stability through Rap1/Rac1 activationScientific Reports 2024 14:12024ISSN 2045--2322
    Chen, Kuan Ju et al.Somatic A-to-I RNA-edited RHOA isoform 2 specific-R176G mutation promotes tumor progression in lung adenocarcinomaMolecular Carcinogenesis2023ISSN 1098--2744
    Zhang, Xingyu et al.Soluble TREM2 ameliorates tau phosphorylation and cognitive deficits through activating transgelin-2 in Alzheimer’s diseaseNature Communications 2023 14:12023ISSN 2041--1723
    Chen, Kuan Ju et al.Somatic A-to-I RNA-edited RHOA isoform 2 specific-R176G mutation promotes tumor progression in lung adenocarcinomaMolecular Carcinogenesis2023ISSN 1098--2744
    Lachowski, Dariusz et al.Substrate Stiffness-Driven Membrane Tension Modulates Vesicular Trafficking via Caveolin-1ACS Nano2022ISSN 1936-086X
    Loria, Rossella et al.SEMA6A/RhoA/YAP axis mediates tumor-stroma interactions and prevents response to dual BRAF/MEK inhibition in BRAF-mutant melanomaJournal of Experimental and Clinical Cancer Research2022ISSN 1756-9966
    Bouzid, Tasneem et al.Rho/ROCK mechanosensor in adipocyte stiffness and traction force generationBiochemical and Biophysical Research Communications2022
    Moztarzadeh, Sina et al.Lack of adducin impairs the stability of endothelial adherens and tight junctions and may be required for cAMP-Rac1-mediated endothelial barrier stabilizationScientific Reports 2022 12:12022ISSN 2045--2322
    Jozic, Ivan et al.Glucocorticoid-mediated induction of caveolin-1 disrupts cytoskeletal organization, inhibits cell migration and re-epithelialization of non-healing woundsCommunications Biology2021ISSN 2399-3642
    Esfahani, Amir Monemian et al.Characterization of the strain-rate–dependent mechanical response of single cell–cell junctionsProceedings of the National Academy of Sciences of the United States of America2021ISSN 1091-6490
    Jin, Xiaowei et al.Modulation of Mechanical Stress Mitigates Anti-Dsg3 Antibody-Induced Dissociation of Cell–Cell AdhesionAdvanced Biology2021ISSN 2701-0198
    Broussard, Joshua A. et al.Desmosomes polarize and integrate chemical and mechanical signaling to govern epidermal tissue form and functionCurrent Biology2021ISSN 1879-0445
    Shoda, Tetsuo et al.Desmoplakin and periplakin genetically and functionally contribute to eosinophilic esophagitisNature Communications2021ISSN 2041-1723
    Bendrick, Jacqueline L. et al.Desmoplakin Harnesses Rho GTPase and p38 Mitogen-Activated Protein Kinase Signaling to Coordinate Cellular MigrationJournal of Investigative Dermatology2019ISSN 1523-1747
    Islam, Rokibul et al.Insulin induces phosphorylation of pyruvate dehydrogenase through RhoA activation pathway in HepG2 cellsFASEB Journal2019ISSN 1530-6860
    Juettner, Vanessa V. et al.VE-PTP stabilizes VE-cadherin junctions and the endothelial barrier via a phosphatase-independent mechanismJournal of Cell Biology2019ISSN 1540-8140
    Richter, Melanie et al.Altered TAOK2 activity causes autism-related neurodevelopmental and cognitive abnormalities through RhoA signalingMolecular Psychiatry2019ISSN 1476-5578
    Tanaka, Leonardo Y. et al.Peri/epicellular protein disulfide isomerase-A1 acts as an upstream organizer of cytoskeletal mechanoadaptation in vascular smooth muscle cellsAmerican Journal of Physiology - Heart and Circulatory Physiology2019ISSN 1522-1539
    Griesi-Oliveira, Karina et al.Actin cytoskeleton dynamics in stem cells from autistic individualsScientific Reports2018ISSN 2045-2322
    Pietrovito, Laura et al.Bone marrow-derived mesenchymal stem cells promote invasiveness and transendothelial migration of osteosarcoma cells via a mesenchymal to amoeboid transitionMolecular Oncology2018ISSN 1878-0261
    Comito, Giuseppina et al.Zoledronic acid impairs stromal reactivity by inhibiting M2-macrophages polarization and prostate cancer-associated fibroblastsOncotarget2017ISSN 1949-2553
    Broussard, Joshua A. et al.The desmoplakin-intermediate filament linkage regulates cell mechanicsMolecular Biology of the Cell2017ISSN 1939-4586
    Haemmerle, Monika et al.Platelets reduce anoikis and promote metastasis by activating YAP1 signalingNature Communications2017ISSN 2041-1723
    Chang, Ching Dong et al.Ursolic acid suppresses hepatitis b virus x protein-mediated autophagy and chemotherapeutic drug resistanceAnticancer Research2016ISSN 1791-7530
    Murugesan, Sricharan et al.Formin-generated actomyosin arcs propel t cell receptor microcluster movement at the immune synapseJournal of Cell Biology2016ISSN 1540-8140
    Fogli, Stefano et al.Salbutamol inhibits RhoA activation in normal but not in desensitized bronchial smooth muscle cellsJournal of Pharmacy and Pharmacology2015ISSN 2042-7158
    Haque, Amranul et al.An engineered N-cadherin substrate for differentiation, survival, and selection of pluripotent stem cell-derived neural progenitorsPLoS ONE2015ISSN 1932-6203
    Justus, Calvin R. et al.GPR4 decreases B16F10 melanoma cell spreading and regulates focal adhesion dynamics through the G13/Rho signaling pathwayExperimental Cell Research2015ISSN 1090-2422
    Luo, Jixian et al.8-Oxoguanine DNA glycosylase-1-mediated DNA repair is associated with Rho GTPase activation and α-smooth muscle actin polymerizationFree Radical Biology and Medicine2014ISSN 1873-4596
    Singh, Jagmohan et al.Aging-associated oxidative stress leads to decrease in IAS tone via RhoA/ROCK downregulationAmerican Journal of Physiology - Gastrointestinal and Liver Physiology2014ISSN 1522-1547
    Mattias, Leino et al.The effects of artificial E-cadherin matrix-induced embryonic stem cell scattering on paxillin and RhoA activation via α-cateninBiomaterials2014ISSN 1878-5905
    Suen, J. Y. et al.Pathway-selective antagonism of proteinase activated receptor 2British Journal of Pharmacology2014ISSN 1476-5381
    Vertelov, Grigory et al.High targeted migration of human mesenchymal stem cells grown in hypoxia is associated with enhanced activation of RhoAStem cell research & therapy2013ISSN 1757-6512
    Schulz, Alexander et al.Merlin isoform 2 in neurofibromatosis type 2-associated polyneuropathyNature Neuroscience2013ISSN 1097-6256
    Kshitiz et al.Matrix rigidity controls endothelial differentiation and morphogenesis of cardiac precursorsScience Signaling2012ISSN 1945-0877
    Kim, Jae Gyu et al.Ras-related GTPases Rap1 and RhoA collectively induce the phagocytosis of serum-opsonized zymosan particles in macrophagesJournal of Biological Chemistry2012ISSN 0021-9258
    Ku, Chin Jen et al.Network crosstalk dynamically changes during neutrophil polarizationCell2012ISSN 1097-4172
    Brown, Jacquelyn A. et al.Neurofibromatosis-1 heterozygosity impairs CNS neuronal morphology in a cAMP/PKA/ROCK-dependent mannerMolecular and Cellular Neuroscience2012ISSN 1044-7431
    Aguilar, Hector N. et al.Phos-tag-based analysis of myosin regulatory light chain phosphorylation in human uterine myocytesPLoS ONE2011ISSN 1932-6203
    Winzeler, Alissa M. et al.The lipid sulfatide is a novel myelin-associated inhibitor of CNS axon outgrowthJournal of Neuroscience2011ISSN 0270-6474

    Question 1:  What is the chemical nature of the Rho activator CN01?

    Answer 1:  CN01 is calpeptin and has the peptide sequence of Z-Leu-Nle-CHO.  The molecular weight is 362.5 and the CAS number is 117591-20-5.  Calpeptin is supplied as a lyophilized white solid and each vial contains 1 mg (10 units) of CN01.  The purity is ≥95% as determined by HPLC.


    Question 2:  Does CN01 selectively activate Rho?

    Answer 2:  No, CN01 (calpeptin) does not specifically activate Rho.  Calpeptin-induced Rho activation is indirect via a mechanism involving inhibition of Shp-2 phosphatase.  Inhibition of Shp-2 allows constitutive activation of Rho GEFs.  CN01 also inhibits calpain-1 in a mechanism that is unrelated to Rho activation.  For a direct and specific Rho activator, please see Cat. # CN03.


    If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com.