Rhosin hydrochloride

Manufactured by Bio-Techne

Sourced in United Kingdom

Rhosin hydrochloride is a chemical compound produced by Bio-Techne for use in laboratory research. It acts as a Rho-associated protein kinase (ROCK) inhibitor. The core function of Rhosin hydrochloride is to inhibit the activity of ROCK proteins, which are involved in regulating cellular processes such as cell proliferation, migration, and contractility.

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Rhosin (8 citations)

5 protocols using rhosin hydrochloride

Actin and Microtubule Modulation Assay

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CK-666 (200μM, 1h; Sigma, SML0006), Latrunculin A (1 μM, 0–2h; Sigma, L5163), Cytochalasin D (100nM, 1–2h; Sigma, C2618), DMSO (1h; Sigma, D2650), Nocodazole (10/25 μM; Sigma, M1404). EHT1864 (20μM, 1h; Tocris), Rhosin hydrochloride (100μM, 1h; Tocris), C3 Transferase (2μg/mL, 3h; Cytoskeleton), Y27632 (10μM, 1h; Sigma), Blebbistatin (85μM, 1h; Sigma), ML141 (25μM, 1h; Millipore Sigma), Wiskostatin (10μM, 1h; TOCRIS), SMIFH2 (25 mM, 1h; Sigma), U0126 (20 μM, 1hr; 19–147, EMD Millipore), Erlotinib (1μg/ml, 1hr; SML2156, Sigma), DMSO (1h; Sigma).

Moore A.S., Coscia S.M., Simpson C.L., Ortega F.E., Wait E.C., Heddleston J.M., Nirschl J.J., Obara C.J., Guedes-Dias P., Boecker C.A., Chew T.L., Theriot J.A., Lippincott-Schwartz J, & Holzbaur E.L. (2021). Actin cables and comet tails organize mitochondrial networks in mitosis. Nature, 591(7851), 659-664.

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Investigating Cell Signaling and Inhibition

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Cells were stimulated or treated with different inhibitors as indicated in the figures. As stimuli, the complement C5a (20 nm; R&D) as well as purified S100A8/S100A8‐homodimer (250 ng mL⁻¹), S100A8/S100A9‐heterodimer (250 ng mL⁻¹), and the mutated S100A8/S100A9‐N70A‐heterodimer (250 ng mL⁻¹) were used. For the inhibition of the Rho GTPases, RhoA, Rac, and Cdc42, the cells were treated with the specific GEF inhibitors Rhosin hydrochloride (30 µm), W56 (250 µm), and ZCL278 (50 µm) for 30 min, respectively (Tocris). Cryptotanshinone was used to inhibit the phosphorylation of STAT3 at a concentration of 5 µm for 30 min (Sigma–Aldrich).

Russo A., Schürmann H., Brandt M., Scholz K., Matos A.L., Grill D., Revenstorff J., Rembrink M., von Wulffen M., Fischer‐Riepe L., Hanley P.J., Häcker H., Prünster M., Sánchez‐Madrid F., Hermann S., Klotz L., Gerke V., Betz T., Vogl T, & Roth J. (2022). Alarming and Calming: Opposing Roles of S100A8/S100A9 Dimers and Tetramers on Monocytes. Advanced Science, 9(36), 2201505.

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Actin and Microtubule Modulation Assay

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Platelet Activation Signaling Pathway Analysis

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Sepharose CL‐2B column, bovine thrombin, and Ca²⁺‐ionophore calcimycin were from Sigma‐Aldrich. CVX was the product of Pentapharm. Phycoerythrin (PE)‐labeled anti‐CD62 antibody (Ab), PE‐cyanine5 (PECy5)‐conjugated anti‐CD41a Ab, PE‐labeled mouse IgG₁, fluorescein isothiocyanate (FITC)‐labeled mouse IgG_2a, PE‐conjugated annexin V, and annexin V binding buffer were purchased from Becton Dickinson. Mouse anti‐human CD41a Ab, Alexa‐fluor 568 annexin V conjugate, and DyLight 405‐labeled goat anti‐mouse Ab were the products of Thermo Fisher Scientific. DyLight 488‐labeled horse anti‐rabbit Ab, Vectashield^® antifade mounting medium, and normal goat serum were from Vector Laboratories. Goat anti‐rabbit IgG conjugated to 15 nm gold particles and goat anti‐mouse IgG conjugated to 10 nm gold particles were purchased from BBI Solutions. Uranyl acetate was from Electron Microscopy Sciences. Lead citrate was made from lead nitrate (VWR International Ltd.). Rabbit anti‐human FXIII‐A Ab and FITC‐labeled mouse anti‐human‐FXIII‐A Ab were produced in our laboratories.¹⁶, ²² The fluorescent calcium indicator Fluo‐4‐AM was obtained from Thermo Fisher Scientific. The RhoA inhibitor Rhosin hydrochloride was purchased from Tocris Bioscience and the transglutaminase inhibitor T101 from Zedira.

Somodi L., Beke Debreceni I., Kis G., Cozzolino M., Kappelmayer J., Antal M., Panyi G., Bárdos H., Mutch N.J, & Muszbek L. (2022). Activation mechanism dependent surface exposure of cellular factor XIII on activated platelets and platelet microparticles. Journal of Thrombosis and Haemostasis, 20(5), 1223-1235.

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Rho GTPase Inhibitor Impacts EV Release

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For mechanistic study, B16F1 or 3T3 Swiss Albino cells were seeded onto 150-mm dishes at a density of 4 or 6 × 10⁶ cells/dish, respectively. After washing the cells with PBS followed by replacing with Advanced DMEM supplemented with 2 mM l-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin, a Rho GTPase inhibitor Rhosin hydrochloride (Tocris Bioscience, Bristol, UK) dissolved in dimethyl sulfoxide (DMSO; Wako Pure Chemical, Osaka, Japan) was added into the culture media to become a concentration of 10 or 30 μM. The final concentration of DMSO in the media was 0.1% in each group. Then, the cells were treated with a constant current of 0.34 mA/cm² for 60 min in the presence or absence of the Rho GTPase inhibitor. After 24 h of additional incubation, the conditioned medium was collected, and EVs in the medium were purified as mentioned above. Thereafter, the number of particles in the collected EV suspension was measured with the nanoparticle multi-analyzer qNano.

Fukuta T., Nishikawa A, & Kogure K. (2019). Low level electricity increases the secretion of extracellular vesicles from cultured cells. Biochemistry and Biophysics Reports, 21, 100713.

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