Product Uses Include
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.
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|Chen, Kuan Ju et al.||Somatic A-to-I RNA-edited RHOA isoform 2 specific-R176G mutation promotes tumor progression in lung adenocarcinoma||Molecular Carcinogenesis||2023||ISSN 1098--2744|
|Lachowski, Dariusz et al.||Substrate Stiffness-Driven Membrane Tension Modulates Vesicular Trafficking via Caveolin-1||ACS Nano||2022||ISSN 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 melanoma||Journal of Experimental and Clinical Cancer Research||2022||ISSN 1756-9966|
|Bouzid, Tasneem et al.||Rho/ROCK mechanosensor in adipocyte stiffness and traction force generation||Biochemical and Biophysical Research Communications||2022|
|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 stabilization||Scientific Reports 2022 12:1||2022||ISSN 2045--2322|
|Jin, Xiaowei et al.||Modulation of Mechanical Stress Mitigates Anti-Dsg3 Antibody-Induced Dissociation of Cell–Cell Adhesion||Advanced Biology||2021||ISSN 2701-0198|
|Broussard, Joshua A. et al.||Desmosomes polarize and integrate chemical and mechanical signaling to govern epidermal tissue form and function||Current Biology||2021||ISSN 1879-0445|
|Esfahani, Amir Monemian et al.||Characterization of the strain-rate–dependent mechanical response of single cell–cell junctions||Proceedings of the National Academy of Sciences of the United States of America||2021||ISSN 1091-6490|
|Jozic, Ivan et al.||Glucocorticoid-mediated induction of caveolin-1 disrupts cytoskeletal organization, inhibits cell migration and re-epithelialization of non-healing wounds||Communications Biology||2021||ISSN 2399-3642|
|Shoda, Tetsuo et al.||Desmoplakin and periplakin genetically and functionally contribute to eosinophilic esophagitis||Nature Communications||2021||ISSN 2041-1723|
|Tanaka, Leonardo Y. et al.||Peri/epicellular protein disulfide isomerase-A1 acts as an upstream organizer of cytoskeletal mechanoadaptation in vascular smooth muscle cells||American Journal of Physiology - Heart and Circulatory Physiology||2019||ISSN 1522-1539|
|Bendrick, Jacqueline L. et al.||Desmoplakin Harnesses Rho GTPase and p38 Mitogen-Activated Protein Kinase Signaling to Coordinate Cellular Migration||Journal of Investigative Dermatology||2019||ISSN 1523-1747|
|Juettner, Vanessa V. et al.||VE-PTP stabilizes VE-cadherin junctions and the endothelial barrier via a phosphatase-independent mechanism||Journal of Cell Biology||2019||ISSN 1540-8140|
|Islam, Rokibul et al.||Insulin induces phosphorylation of pyruvate dehydrogenase through RhoA activation pathway in HepG2 cells||FASEB Journal||2019||ISSN 1530-6860|
|Richter, Melanie et al.||Altered TAOK2 activity causes autism-related neurodevelopmental and cognitive abnormalities through RhoA signaling||Molecular Psychiatry||2019||ISSN 1476-5578|
|Griesi-Oliveira, Karina et al.||Actin cytoskeleton dynamics in stem cells from autistic individuals||Scientific Reports||2018||ISSN 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 transition||Molecular Oncology||2018||ISSN 1878-0261|
|Broussard, Joshua A. et al.||The desmoplakin-intermediate filament linkage regulates cell mechanics||Molecular Biology of the Cell||2017||ISSN 1939-4586|
|Haemmerle, Monika et al.||Platelets reduce anoikis and promote metastasis by activating YAP1 signaling||Nature Communications||2017||ISSN 2041-1723|
|Comito, Giuseppina et al.||Zoledronic acid impairs stromal reactivity by inhibiting M2-macrophages polarization and prostate cancer-associated fibroblasts||Oncotarget||2017||ISSN 1949-2553|
|Chang, Ching Dong et al.||Ursolic acid suppresses hepatitis b virus x protein-mediated autophagy and chemotherapeutic drug resistance||Anticancer Research||2016||ISSN 1791-7530|
|Murugesan, Sricharan et al.||Formin-generated actomyosin arcs propel t cell receptor microcluster movement at the immune synapse||Journal of Cell Biology||2016||ISSN 1540-8140|
|Haque, Amranul et al.||An engineered N-cadherin substrate for differentiation, survival, and selection of pluripotent stem cell-derived neural progenitors||PLoS ONE||2015||ISSN 1932-6203|
|Justus, Calvin R. et al.||GPR4 decreases B16F10 melanoma cell spreading and regulates focal adhesion dynamics through the G13/Rho signaling pathway||Experimental Cell Research||2015||ISSN 1090-2422|
|Fogli, Stefano et al.||Salbutamol inhibits RhoA activation in normal but not in desensitized bronchial smooth muscle cells||Journal of Pharmacy and Pharmacology||2015||ISSN 2042-7158|
|Singh, Jagmohan et al.||Aging-associated oxidative stress leads to decrease in IAS tone via RhoA/ROCK downregulation||American Journal of Physiology - Gastrointestinal and Liver Physiology||2014||ISSN 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-catenin||Biomaterials||2014||ISSN 1878-5905|
|Suen, J. Y. et al.||Pathway-selective antagonism of proteinase activated receptor 2||British Journal of Pharmacology||2014||ISSN 1476-5381|
|Luo, Jixian et al.||8-Oxoguanine DNA glycosylase-1-mediated DNA repair is associated with Rho GTPase activation and α-smooth muscle actin polymerization||Free Radical Biology and Medicine||2014||ISSN 1873-4596|
|Vertelov, Grigory et al.||High targeted migration of human mesenchymal stem cells grown in hypoxia is associated with enhanced activation of RhoA||Stem cell research & therapy||2013||ISSN 1757-6512|
|Schulz, Alexander et al.||Merlin isoform 2 in neurofibromatosis type 2-associated polyneuropathy||Nature Neuroscience||2013||ISSN 1097-6256|
|Brown, Jacquelyn A. et al.||Neurofibromatosis-1 heterozygosity impairs CNS neuronal morphology in a cAMP/PKA/ROCK-dependent manner||Molecular and Cellular Neuroscience||2012||ISSN 1044-7431|
|Ku, Chin Jen et al.||Network crosstalk dynamically changes during neutrophil polarization||Cell||2012||ISSN 1097-4172|
|Kshitiz et al.||Matrix rigidity controls endothelial differentiation and morphogenesis of cardiac precursors||Science Signaling||2012||ISSN 1945-0877|
|Kim, Jae Gyu et al.||Ras-related GTPases Rap1 and RhoA collectively induce the phagocytosis of serum-opsonized zymosan particles in macrophages||Journal of Biological Chemistry||2012||ISSN 0021-9258|
|Aguilar, Hector N. et al.||Phos-tag-based analysis of myosin regulatory light chain phosphorylation in human uterine myocytes||PLoS ONE||2011||ISSN 1932-6203|
|Winzeler, Alissa M. et al.||The lipid sulfatide is a novel myelin-associated inhibitor of CNS axon outgrowth||Journal of Neuroscience||2011||ISSN 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.
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