rabbit polyclonal anti phospho plk1  (Cell Signaling Technology Inc)

Journal: Cell reports

Article Title: Feedback control of PLK1 by Apolo1 ensures accurate chromosome segregation

doi: 10.1016/j.celrep.2021.109343

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-pT210-PLK1 , Cell Signaling Technology , Cat#9062; RRID:AB_11127447.

Techniques: Virus, Recombinant, Activity Assay, Software

Journal: STAR Protocols

Article Title: Protocol to identify centrosome-associated transcription factors during mitosis in mammalian cell lines

doi: 10.1016/j.xpro.2021.100495

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-phospho-PLK1 (Thr210) (reactivity: human) , Cell Signaling Technology , 5472S.

Techniques: Recombinant, Protease Inhibitor, Plasmid Preparation, Bicinchoninic Acid Protein Assay, In Situ, Software, Membrane, Laser-Scanning Microscopy

Journal: Acta Pharmaceutica Sinica. B

Article Title: MCC1019, a selective inhibitor of the Polo-box domain of Polo-like kinase 1 as novel, potent anticancer candidate

doi: 10.1016/j.apsb.2019.02.001

Figure Lengend Snippet: Inhibition of PLK1 PBD by MCC1019. (A) Inhibition of the binding of fluorescein-labeled phosphopeptides to PLK1, PLK2 or PLK3 PBDs by MCC1019 using a fluorescence polarization assay. The data are represented as mean ± SD of three independent experiments. (B) Thermophoresis binding curve of MCC1019 to the PLK1 and PLK1 PBD obtained by microscale thermopheresis. (C) Surface visualization of molecular docking of MCC1019 to the PLK1 PBD (PDB: 4X9R). (D) SeeSAR visualization of MCC1019 binding to human PLK1 (PDB: 4X9R): HYDE corona coloring is based on atomic affinity: green for favourable and red for unfavourable predicted affinities between atoms and amino acids. (E) Dose response curves of MCC1019 for different cancer cell lines obtained by cytotoxicity resazurin reduction assay, with westernblot analysis of PLK1 expression on each cell. The data are represented as mean ± SD of three independent experiments.

Article Snippet: Then, membranes were incubated with primary antibodies [anti-BUBR1 rabbit polyclonal antibody (1:1000, Abcam, Cambridge, UK), anti-phospho-AKT (Ser473) antibody (1:1000, Cell Signaling, Frankfurt, Germany), anti-phospho-FOXO1(Ser256) antibody (1:1000, Cell Signaling), anti-HIF-1 α antibody (1:1000, Cell Signaling), anti-PTEN antibody (1:1000, Cell Signaling), anti-PARP polyclonal rabbit antibody (1:1000, Cell Signaling,), anti-LC3B polyclonal rabbit antibody (1:1000, Cell Signaling), anti-beclin-1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-PLK1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-phospho-PLK1 (Thr210) polyclonal rabbit antibody (1:1000, Cell Signaling), anti-cyclin B1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-phospho-histone H 3 (Ser10) polyclonal rabbit antibody (1:1000, Cell Signaling), or anti- β -actin polyclonal rabbit antibody (1:2000, Cell Signaling)] overnight at 4 °C.

Techniques: Inhibition, Binding Assay, Labeling, Fluorescence, Expressing

Journal: Acta Pharmaceutica Sinica. B

Article Title: MCC1019, a selective inhibitor of the Polo-box domain of Polo-like kinase 1 as novel, potent anticancer candidate

doi: 10.1016/j.apsb.2019.02.001

Figure Lengend Snippet: MCC1019 inhibit downstream effector proteins of PLK1. (A) Western blot analysis of BUBR1, p-AKT, p-FOXO1 and HIF-1 α after treatment with different concentrations of MCC1019 for 24 h. Data represent relative expression intensity to β -actin, Error bars are ± SD of three independent experiments. (B) Diagram showing molecular effects of PLK1 inhibition on AKT signaling pathway that appeared in the study.

Article Snippet: Then, membranes were incubated with primary antibodies [anti-BUBR1 rabbit polyclonal antibody (1:1000, Abcam, Cambridge, UK), anti-phospho-AKT (Ser473) antibody (1:1000, Cell Signaling, Frankfurt, Germany), anti-phospho-FOXO1(Ser256) antibody (1:1000, Cell Signaling), anti-HIF-1 α antibody (1:1000, Cell Signaling), anti-PTEN antibody (1:1000, Cell Signaling), anti-PARP polyclonal rabbit antibody (1:1000, Cell Signaling,), anti-LC3B polyclonal rabbit antibody (1:1000, Cell Signaling), anti-beclin-1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-PLK1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-phospho-PLK1 (Thr210) polyclonal rabbit antibody (1:1000, Cell Signaling), anti-cyclin B1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-phospho-histone H 3 (Ser10) polyclonal rabbit antibody (1:1000, Cell Signaling), or anti- β -actin polyclonal rabbit antibody (1:2000, Cell Signaling)] overnight at 4 °C.

Techniques: Western Blot, Expressing, Inhibition

Journal: Acta Pharmaceutica Sinica. B

Article Title: MCC1019, a selective inhibitor of the Polo-box domain of Polo-like kinase 1 as novel, potent anticancer candidate

doi: 10.1016/j.apsb.2019.02.001

Figure Lengend Snippet: Induction of mitotic arrest by MCC1019. (A) Flow cytometric cell cycle analysis of exponentially growing A549 cells treated with MCC1019 for 24 h in a concentration range from 10 to 40 μmol/L. The data are represented as mean ± SD of three independent experiments. (B) Western blot analysis of A549 cell lysates treated with MCC1019 for 8, 16, 20, 24, 48 or 72 h. Increased expression levels of PLK1 and cyclin B1 were seen after 24 h treatment. (C) Western blot analysis of the mitotic markers PLK1, cyclin B1 and p-HH3 after treatment with different concentrations of MCC1019 for 24 h. Data represent relative expression intensity to β- actin, Error bars are ± SD of three independent experiments. (D) Immunofluorescence analysis of A549 treated with MCC1019 and DMSO (control) and stained for α -tubulin (green) and DNA (blue). The data are represented as mean ± SD of cells undergoing mitosis.

Article Snippet: Then, membranes were incubated with primary antibodies [anti-BUBR1 rabbit polyclonal antibody (1:1000, Abcam, Cambridge, UK), anti-phospho-AKT (Ser473) antibody (1:1000, Cell Signaling, Frankfurt, Germany), anti-phospho-FOXO1(Ser256) antibody (1:1000, Cell Signaling), anti-HIF-1 α antibody (1:1000, Cell Signaling), anti-PTEN antibody (1:1000, Cell Signaling), anti-PARP polyclonal rabbit antibody (1:1000, Cell Signaling,), anti-LC3B polyclonal rabbit antibody (1:1000, Cell Signaling), anti-beclin-1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-PLK1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-phospho-PLK1 (Thr210) polyclonal rabbit antibody (1:1000, Cell Signaling), anti-cyclin B1 polyclonal rabbit antibody (1:1000, Cell Signaling), anti-phospho-histone H 3 (Ser10) polyclonal rabbit antibody (1:1000, Cell Signaling), or anti- β -actin polyclonal rabbit antibody (1:2000, Cell Signaling)] overnight at 4 °C.

Techniques: Cell Cycle Assay, Concentration Assay, Western Blot, Expressing, Immunofluorescence, Control, Staining