Bi2536

Manufactured by Thermo Fisher Scientific

BI2536 is a small molecule that functions as a potent and selective inhibitor of the serine/threonine-protein kinase PLK1 (Polo-like kinase 1). It is commonly used in research applications as a tool compound to study cellular processes related to the regulation of mitosis and cell cycle progression.

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Lab products found in correlation

Az3146, by Bio-Techne (2 mentions) Nuclease free water, by New England Biolabs (1 mentions) Binucleine 2, by Merck Group (1 mentions) Cytochalasin d, by Merck Group (1 mentions) Taxol, by Thermo Fisher Scientific (1 mentions) Flavopiridol, by Santa Cruz Biotechnology (1 mentions) Nocodazole, by Merck Group (1 mentions) Nsc697923, by Merck Group (1 mentions) Zm 447439, by R&D Systems (1 mentions) Pr 619, by Merck Group (1 mentions) Vx 680, by LC Laboratories (1 mentions) Flavopiridol, by Merck Group (1 mentions) Co2 independent media, by Thermo Fisher Scientific (1 mentions) 3 mb pp1, by Merck Group (1 mentions) Thymidine, by Merck Group (1 mentions)

3 protocols using bi2536

Drosophila Embryonic Microtubule Manipulation

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Embryos were staged according to criteria established by Campos-Ortega and Hartenstein [32 (link)]. Embryos were collected on grape-juice agar plates, aged on collection plates and dechorionated by hand. Dechorionated embryos were briefly desiccated and microinjected as previously described [33 (link)]. Needle concentrations for injected solutions were as follows: Colchicine and paclitaxel (Sigma-Aldrich) [5μM], Cytochalasin D (Sigma-Aldrich), Binucleine 2 (Sigma-Aldrich), and BI 2536 (Thermo Fisher) [10μM], Ciliobrevin D (Calbiochem) [100μM]. All drugs were dissolved in DMSO which was limited to a final concentration of 10%. Tubulin HiLyte 488 and 647 (Cytoskeleton Inc.) was injected at 1mg/ml in nuclease free water (New England Biolabs). Microinjection of embryos was performed with ~0.1 μl of RTNL1 dsRNA (1.5μg/μl) injected into the dorsoanterior region of the embryo during the syncytial blastoderm stage.

Diaz U., Bergman Z.J., Johnson B.M., Edington A.R., de Cruz M.A., Marshall W.F, & Riggs B. (2019). Microtubules are necessary for proper Reticulon localization during mitosis. PLoS ONE, 14(12), e0226327.

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Assessing Small Molecule Inhibitors

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All chemicals were resuspended in DMSO. NSC697923 (Sigma-Aldrich) was used at 20 µM. Nocodazole (Sigma-Aldrich) was used at 50 nM. MG132 (EMD Biosciences) was used at 10 µM. CC0651 (Thermo Fisher Scientific) was used at 50 µM. Taxol (Life Technologies) was used at 1 µM. PYR-41 (Santa Cruz Biotechnology) was used at 20 µM. TAK-243 (formerly MLN7243) was a gift from Hidde Ploegh (Whitehead Institute) and used at 25 µM. PR-619 (Sigma-Aldrich) was used at 100 µM. ZM447439 (R&D Systems) was used at 2 µM. VX680 (LC Laboratories) was used at 2.5 µM. Flavopiridol (Santa Cruz Biotechnology) was used at 5 µM. AZ3146 (Tocris) was used at 2 µM. BI2536 (Thermo Fisher Scientific) was used at 10 µM. Okadaic acid (VWR) was used at 1 µM. To dose cells, drugs were diluted in fresh media and then added to the cells. For immunofluorescence, cells were dosed for 15 min before fixation.

Monda J.K, & Cheeseman I.M. (2018). Dynamic regulation of dynein localization revealed by small molecule inhibitors of ubiquitination enzymes. Open Biology, 8(9), 180095.

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Culturing and Treatment of Cell Lines

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HeLa cell lines were cultured as described previously in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), penicillin/streptomycin and 2 mM L-glutamine. hTERT-RPE1 Plk1^as cells were maintained in DMEM:F12 with 10% fetal bovine serum (FBS), penicillin/streptomycin and 2 mM L-glutamine. For time-lapse imaging, cells were maintained in CO₂-independent media (Invitrogen) with 10% FBS.
Unless otherwise indicated, cells were incubated in 10 μM BI2536 (Thermo Fisher Scientific) for 2.5 h, although severely defective CENP-A deposition was observed at concentrations down to at least 10 nM (Fig. S2A). Where indicated, cells were incubated with 5 μM flavopiridol (Sigma), 2 μM AZ3146 (Tocris), 10 μM 3MB-PP1 (Merck), or 10 μM STLC (Sigma) for 1-2.5 h. HeLa cells were synchronized by double thymidine block using 2 mM thymidine (Sigma) for all immunofluorescence and live cell imaging experiments unless otherwise stated.

McKinley K.L, & Cheeseman I.M. (2014). Polo-like kinase 1 licenses CENP-A deposition at centromeres. Cell, 158(2), 397-411.

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