Journal: The EMBO Journal

Article Title: Requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1038/s44318-024-00228-9

Figure Lengend Snippet: ( A ) Pericentromeric cohesin is deprotected long before sister chromatids separate. During phosphorylation- and Wapl-dependent release of cohesin from chromosome arms, pericentromeric cohesin is protected by associated Sgo1-PP2A. Following the early mitotic degradation of Nek2a and cyclin A2, Sgo1-PP2A relocates to kinetochores. Yet, pericentromeric cohesion persists until securin and cyclin B1 are degraded and separase is activated. A requirement for Nek2a and cyclin A2 for prophase pathway signaling would explain why, in metaphase, Wapl can no longer release cohesin. ( B ) Nek2a and Cdk1/2-cyclin A2 are necessary for Wapl-dependent in vitro displacement of cohesin from chromatin. Chromatin was freshly purified by repeated sedimentation from G2-arrested HeLaK cells, incubated with recombinant Wapl, kinases, and inhibitors (BI2536, ZM-447439, and RO-3306), as indicated, and then re-pelleted. Supernatants and chromatin pellets were analyzed by immunoblotting. ( C ) Wapl and the three kinases Nek2a, Cdk1/2-cyclin A2, and Aurora B-INCENP are sufficient to support the displacement of cohesin from isolated chromatin. Chromatin was freshly purified by repeated sedimentation from G2-arrested HeLaK cells, incubated with recombinant Wapl, Nek2a, Cdk1/2-cyclin A2, Cdk1-cyclin B1, aurora B, and/or Plk1, as indicated, and then re-pelleted. Supernatants and chromatin pellets were analyzed by immunoblotting. .

Article Snippet: Recombinant human active PLK1 Protein , R&D Systems , 3804-KS-010.

Techniques: In Vitro, Purification, Sedimentation, Incubation, Recombinant, Western Blot, Isolation

Journal: The EMBO Journal

Article Title: Requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1038/s44318-024-00228-9

Figure Lengend Snippet: ( A ) Recombinant kinases are active as judged by the phosphorylation of model substrates. Plk1, aurora B-INCENP, Nek2a, Cdk1/2-cyclin A2, or Cdk1-cyclin B1 supplemented with their specific inhibitor or carrier solvent DMSO (−) were incubated with the corresponding model substrate in the presence of [γ 33 P]-ATP. Reactions were subjected to SDS-PAGE followed by autoradiography. BI BI2536, ZM ZM-447439, ST staurosporine, RO RO-3306, MBP myelin basic protein. ( B ) Recombinant Nek2a, Cdk1/2-cyclin A2, and Wapl are free of cyclin B1 and Plk1. The preparations of Nek2a, Cdk1/2-cyclin A2, Wapl, Cdk1-cyclin B1, and Plk1 used for the release assays were characterized by immunoblotting using the indicated antibodies. In vitro expressed (IVTT) cyclin B1 served as an additional control.

Article Snippet: Recombinant human active PLK1 Protein , R&D Systems , 3804-KS-010.

Techniques: Recombinant, Solvent, Incubation, SDS Page, Autoradiography, Western Blot, In Vitro, Control

Journal: The EMBO Journal

Article Title: Requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1038/s44318-024-00228-9

Figure Lengend Snippet: ( A ) Experimental outline. DNA-mediated chromatin pull-down from G2-arrested HeLaK cells is followed by cohesin eviction experiments. ( B ) Cohesin is eluted from immobilized chromatin upon DNA-cleavage but not high salt treatment. Chromatin beads were treated with benzonase and/or high salt, and retained proteins were detected by immunoblotting. High-salt-washed chromatin (lane 6) was used for subsequent cohesin eviction experiments. ( C ) Wapl-dependent cohesin release in vitro requires Nek2a, Cdk1/2-cyclin A2, and Aurora B but not Cdk1-cyclin B1 and Plk1. Immobilized chromatin was combined with the indicated proteins. Following centrifugation, DNA-beads and supernatant were analyzed by immunoblotting. .

Article Snippet: Recombinant human active PLK1 Protein , R&D Systems , 3804-KS-010.

Techniques: Western Blot, In Vitro, Centrifugation

Journal: The EMBO Journal

Article Title: Requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1038/s44318-024-00228-9

Figure Lengend Snippet: ( A ) Nek2a and Cdk1/2-cyclin A2 target juxtaposed but distinct residues within the disordered, C-terminal domain of Pds5b. In vitro expressed Pds5b variants were incubated with the indicated kinases and inhibitors in the presence of [γ- 33 P]-ATP, subjected to SDS-PAGE, and analyzed by autoradiography and immunoblotting. Note that Pds5b is also phosphorylated by aurora B and, weakly, by Plk1. KD kinase-dead Nek2a-Lys37Met; IVTT coupled in vitro transcription-translation. ( B ) Graphical summary of Nek2a- and Cdk1/2-cyclin A2-targeted Pds5b residues and corresponding phosphorylation-resistant variants. ( C ) Ser1209 of Pds5b is phosphorylated by Nek2a but not Cdk1/2-cyclin A2. Following incubation of in vitro expressed Pds5b-WT or −3A with Nek2a or Cdk1/2-cyclin A2, samples were analyzed by immunoblotting using the indicated antibodies. Nek2a-Lys37Met (KD, kinase-dead) served as a negative control. ( D ) The spatiotemporal dynamics of Ser1209-phosphorylation correlate with the recruitment of Wapl to chromatin followed by cohesin release. HeLaK cells synchronously cycling through mitosis were subjected to time-resolved fractionation and immunoblotting. At the time of release from a RO-3306-mediated arrest, cultures were supplemented with a Nek2a inhibitor (NCL00017509) or carrier solvent (mock). .

Article Snippet: Recombinant human active PLK1 Protein , R&D Systems , 3804-KS-010.

Techniques: In Vitro, Incubation, SDS Page, Autoradiography, Western Blot, Negative Control, Fractionation, Solvent

Journal: The EMBO Journal

Article Title: Requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1038/s44318-024-00228-9

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Recombinant human active PLK1 Protein , R&D Systems , 3804-KS-010.

Techniques: Recombinant, Transduction, Sequencing, Protease Inhibitor, Membrane, Software

Journal: bioRxiv

Article Title: In vitro reconstitution reveals requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1101/2024.03.12.584565

Figure Lengend Snippet: A Recombinant kinases are active as judged by phosphorylation of model substrates. Plk1, aurora B-INCENP, Nek2a, Cdk1/2-cyclin A2 or Cdk1-cyclin B1 supplemented with their specific inhibitor or carrier solvent DMSO (-) were incubated with corresponding model substrate in presence of [γ 33 P]-ATP. Reactions were subjected to SDS-PAGE followed by autoradiography. BI, BI2536; ZM, ZM447439; ST, staurosporine; RO, RO3306; MBP, myelin basic protein. B Nek2a and Cdk1/2-cyclin A2 are necessary for Wapl-dependent in vitro displacement of cohesin from chromatin. Chromatin was freshly purified by repeated sedimentation from G2-arrested HeLaK cells, incubated with recombinant Wapl, kinases and inhibitors (BI2536, ZM447439 and RO3306), as indicated, and then re-pelleted. Supernatants and chromatin pellets were analyzed by immunoblotting. C Wapl and the three kinases Nek2a, Cdk1/2-cyclin A2 and Aurora B-INCENP are sufficient to support displacement of cohesin from isolated chromatin. Experiment was done as in B).

Article Snippet: Active Plk1 was purchased from R&D Systems (No. 3804-KS).

Techniques: Recombinant, Phospho-proteomics, Solvent, Incubation, SDS Page, Autoradiography, In Vitro, Purification, Sedimentation, Western Blot, Isolation

Journal: bioRxiv

Article Title: In vitro reconstitution reveals requirement of Nek2a and cyclin A2 for Wapl-dependent removal of cohesin from prophase chromatin

doi: 10.1101/2024.03.12.584565

Figure Lengend Snippet: A Nek2a and Cdk1/2-cyclin A2 target juxtaposed but distinct residues within the disordered, C-terminal domain of Pds5b. In vitro expressed Pds5b variants were incubated with the indicated kinases and inhibitors in presence of [γ- 33 P]-ATP, subjected to SDS-PAGE and analyzed by autoradiography and immunoblotting. Note that Pds5b is also phosphorylated by aurora B and, weakly, by Plk1. KD = kinase-dead Nek2a-Lys37Met; IVTT = coupled in vitro transcription-translation. B Rendering Pds5b resistant against both Nek2a and Cdk1/2-cyclin A2 is necessary and sufficient to switch it from a Wapl- to a sororin-binder. Shown are the experimental outline and corresponding immunoblots. Note that following the incubation with Nek2a and/or Cdk1/2-cyclin A2, ATP was no longer present to exclude phosphorylation of sororin (and/or Wapl) by traces of kinases, which might be retained on the beads.

Article Snippet: Active Plk1 was purchased from R&D Systems (No. 3804-KS).

Techniques: In Vitro, Incubation, SDS Page, Autoradiography, Western Blot, Phospho-proteomics

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: ( A ) Representative time-lapse images of primary MEFs derived from wildtype (top) and Gravin knockout (bottom) cells. Cells transiently expressing GFP-H2B were monitored through mitosis. ( B ) Quantification of time-lapse experiments depicts duration of mitosis from nuclear envelope breakdown to the onset of anaphase; Wildtype, n = 199, Gravin KO, n = 121, ****p<0.0001. ( C, D ) Structured illumination microscopy (SIM) of representative mitotic HEK293 cells stably expressing Control or Gravin shRNA. Composite images (left) depict cells stained for α-tubulin (green) and DNA (blue). Immunofluorescence of pT210-Plk1 ( C ) and pT288-AurA ( D ) as an index of kinase activity (mid) and 5X magnification of centrosomal pT210-Plk1 ( C ) and pT288-AurA ( D ) signals (insets). ( E, F ) Quantification of centrosomal pT210-Plk1 ( E ) and pT288-AurA ( F ) immunofluorescence. Points represent individual cells ( n ). Data are normalized to Con shRNA; ( E ) Con shRNA, n = 45, Gra shRNA, n = 46, **p=0.0036; ( F ) Con shRNA, n = 62, Gra shRNA, n = 66, ****p<0.0001. Experiments were conducted at least three times (N = 3) and P values were calculated by unpaired two-tailed Student’s t-test. Data are mean ± s.e.m. ( G ) SIM micrographs of Gravin (top, gray and magenta) in interphase and pT766-Gravin (bottom, gray and magenta) in mitotic U2OS cells. Composite images (right) also depict α-tubulin (green) and DNA (blue). ( H ) Schematic of global drug distribution (gray) vs drug targeting to centrosomes (green). Gravin scaffolds centrosome-localized pools of Plk1 and AurA.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Derivative Assay, Knock-Out, Expressing, Microscopy, Stable Transfection, Control, shRNA, Staining, Immunofluorescence, Activity Assay, Two Tailed Test

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: ( A ) Schematic of a centrosome-directed LoKI platform. SNAP-PACT fusion proteins conjugate CLP-linked Plk1 inhibitors at centrosomes. Inset depicts BI2536 in the ATP-binding pocket of Plk1. ( B ) Chemical structure of CLP-BI2536. ( C ) Dose-response curve of in vitro Plk1 inhibition with CLP-BI2536. ( D ) Structured illumination microscopy (SIM) of a LoKI-on U2OS cell labeled with CLP-fluorescein. Immunofluorescent detection of α-tubulin (green), DNA (blue), mCherry-SNAP-PACT (magenta) and CLP-fluorescein (yellow). Magnification of SNAP and CLP-fluorescein co-distribution at a centrosome (inset). ( E ) SIM micrographs of LoKI-off (left) and LoKI-on (right) U2OS cells. SNAP expression (top, magenta), CLP-fluorescein conjugation (mid, yellow) and composite images (bottom) are depicted. ( F ) Pulse-chase experiments measuring CLP-BI2536’s ability to block CLP-rhodamine conjugation to LoKI-on. In-gel rhodamine fluorescence (top), immunoblot of SNAP loading controls (mid), and fluorescence quantification of pulse-chase experiments (bottom). ( G, H ) Immunofluorescence of representative mitotic LoKI-off ( G ) and LoKI-on ( H ) U2OS cells treated with DMSO or 250 nM CLP-BI2536 for 4 hr. Composite images (left) show α-tubulin (green), DNA (blue), and SNAP (magenta). Immunofluorescent detection of pT210-Plk1 (mid, gray) as an index of kinase activity. 5X magnification of centrosomal pT210-Plk1 signals and surface plots measuring integrated intensity of pT210-Plk1 signal (insets). ( I, J ) Quantification of centrosomal pT210-Plk1 immunofluorescence for LoKI-expressing cells. Points represent individual cells ( n ). Data normalized to DMSO. Application of DMSO or CLP-BI2536 for 4 hr, ( I ) 100 nM, LoKI-off, n = 46, LoKI-on, n = 59, **p=0.0059; 250 nM, LoKI-off, n = 46, LoKI-on, n = 46, ****p<0.0001 and drug treatment followed by 1 hr washout ( J ) 250 nM, LoKI-off, n = 24, LoKI-on, n = 42, ****p<0.0001. Experiments were conducted at least three times (N = 3) and P values were calculated by unpaired two-tailed Student’s t-test. Data are mean ± s.e.m. NS, not significant. Source files for analysis of pulse-chase experiments are available in and for quantification of pT210-Plk1 are available in . Figure 2—source data 1. Analysis for pulse-chase experiments with CLP-BI2536 in SNAP-PACT cells. Figure 2—source data 2. Raw analysis for pT210-Plk1 signal.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Binding Assay, In Vitro, Inhibition, Microscopy, Labeling, Expressing, Conjugation Assay, Pulse Chase, Blocking Assay, Fluorescence, Western Blot, Immunofluorescence, Activity Assay, Two Tailed Test

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: ( A ) Full chemical structure of CLP-BI2536. ( B ) Dose-response curve depicting in vitro Plk1 inhibition with increasing concentrations of CLP-BI2536 conjugated to purified SNAP. ( C ) Schematic of LoKI viral construct with mCherry-SNAP-PACT under control of a doxycycline-inducible promoter. ( D ) Immunoblot confirming SNAP-PACT (top) expression after induction with doxycycline for 72 hr and GAPDH loading controls (bottom). ( E ) Immunoblot of SNAP-PACT (top) expression at selected time points after removal of doxycycline and GAPDH loading controls (bottom). Quantification of amalgamated data is presented below. ( F ) Immunofluorescent detection of interphase (top) and mitotic (bottom) U2OS cells showing α-tubulin (left and green), DNA (mid and blue), and SNAP (right and magenta). ( G, H ) Diagram of centrosomal LoKI-on ( G ) platform with drugs conjugated and LoKI-off ( H ) platform containing a mutation that occludes CLP binding. Experiments were conducted at least two times (N = 2–3). Data are mean ± s.e.m.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: In Vitro, Inhibition, Purification, Construct, Control, Western Blot, Expressing, Mutagenesis, Binding Assay

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: ( A ) Immunofluorescence detection of pT210-Plk1 as an index of kinase activity in parental U2OS cells treated with DMSO or unconjugated BI2536 for 4 hr. ( B ) Quantification of centrosomal pT210-Plk1 immunofluorescence collected from parental U2OS cells. ( C ) Quantification of total Plk1 immunofluorescence at centrosomes in LoKI-expressing cells after 4 hr CLP-BI2536 treatment; 250 nM, LoKI-off, n = 55, LoKI-on, n = 47, *p=0.0217; 500 nM, LoKI-off, n = 52, LoKI-on, n = 51, *p=0.0295. ( D ) Quantification of pT210-Plk1 immunofluorescence in control cells lacking SNAP expression (not induced with doxycycline) after 4 hr CLP-BI2536 treatment. ( E ) Immunoblot detection of pT210-Plk1 (blot 2) and pT288-AurA (blot 4) in LoKI-off and LoKI-on expressing cells collected via mitotic shake-off. Cells were treated for 16 hr with nocodazole and 4 hr with nocodazole plus DMSO, 250 nM CLP-BI2536, or 100 nM CLP-MLN8237. Total Plk1 (blot 3), AurA (blot 5), SNAP-PACT (blot 1), and GAPDH (blot 6) are also depicted. ( F ) Immunoblot confirming SNAP-PACT (top) expression after induction with doxycycline for 72 hr in RPE and HeLa cells and GAPDH loading controls (bottom). ( G–J ) Immunofluorescence detection of pT210-Plk1 at centrosomes in LoKI-off (top) and LoKI-on (bottom) RPE ( G ) and HeLa ( I ) cells treated with 250 nM CLP-BI2536 for 4 hr. Quantification of pT210-Plk1 immunofluorescence at centrosomes in RPE ( H ) 250 nM, LoKI-off, n = 70, LoKI-on, n = 47, ****p<0.0001 and HeLa ( J ) 250 nM, LoKI-off, n = 42, LoKI-on, n = 46, ****p<0.0001 LoKI-expressing cells after 4 hr CLP-BI2536 treatment. Points represent individual cells ( n ). Data normalized to DMSO. Experiments were conducted at least three times (N = 3) and P values were calculated by unpaired two-tailed Student’s t-test. Data are mean ± s.e.m. NS, not significant.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Immunofluorescence, Activity Assay, Expressing, Control, Western Blot, Two Tailed Test

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: Quantification of non-normalized pT210-Plk1 immunofluorescence signal at centrosomes in U2OS ( A ) RPE ( B ) and HeLa ( C ) LoKI-expressing cells after treatment with indicated concentrations of CLP-BI2536 for 4 hr. Points represent individual cells ( n ). Experiments were conducted at least three times (N = 3). Data are mean ± s.e.m.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Immunofluorescence, Expressing

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: ( A ) Schematic depicting bipolar (top), abnormal bipolar (mid), and monopolar (bottom) mitotic spindle classifications. ( B ) Representative composite (left) images show α-tubulin (green), DNA (blue), and SNAP (magenta) or α-tubulin (right, gray) staining alone for each spindle type in U2OS cells. ( C–E ) Spindle profile measurements of U2OS ( C ) RPE ( D ) and HeLa ( E ) cells treated with 250 nM CLP-BI2536 for 4 hr. Spindle profiling indicates the % of each spindle type in drug-treated LoKI-on and LoKI-off cells. Number of cells analyzed per condition are indicated; ( C ) n = 3, *p=0.0214; ( D ) n = 3, *p=0.0269. ( F ) Schematic depicting that Plk1 phosphorylation of pericentriolar substrates facilitates accumulation of γ-tubulin at centrosomes and microtubule nucleation. ( G, H ) Representative composite (left) images show γ-tubulin (yellow) and DNA (blue) in U2OS cells expressing LoKI-off ( G ) and LoKI-on ( H ) treated with 250 nM CLP-BI2536 for 4 hr, followed by a 1 hr washout. 5X magnified grayscale images of centrosomal γ-tubulin (right). ( I ) Quantification of centrosomal γ-tubulin immunofluorescence in LoKI-expressing cells. Points represent individual cells ( n ). Data normalized to DMSO. A ROUT (Q = 1%) outlier test was performed and two values were removed prior to performing statistical tests; LoKI-off, n = 66, LoKI-on, n = 74, ****p<0.0001. Experiments were conducted at least three times (N = 3) and P values were calculated by unpaired two-tailed Student’s t-test. Data are mean ± s.e.m. NS, not significant. Source files for spindle profile analyses are available in . Figure 3—source data 1. Spindle profile analyses.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Staining, Phospho-proteomics, Expressing, Immunofluorescence, Two Tailed Test

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet: ( A ) Schematic of experimental scheme. Microinjection of LoKI mRNAs into zebrafish embryos occurs at the 1–2 cell stage. Live-cell imaging is conducted at ~50% epiboly. ( B ) Zebrafish embryos (left, brightfield) depicting regional expression of SNAP (mid, magenta) at ~50% epiboly. Composite images (right) depict expression of SNAP only in the cells of the zebrafish embryos. ( C ) Centrosomal delivery of Plk1 inhibitors perturb cell division in zebrafish embryos. Time-lapse images of dividing cells embedded in LoKI-on zebrafish embryos 5 hr post application of 250 nM CLP-BI2536. Representative examples of normal bipolar spindles (top) multipolar spindles (mid) and spindle orientation defects (bottom) are presented. Composite images show microtubule marker EMTB-3xGFP (white) and SNAP (magenta). ( D, E ) 3D-rendered images depict incidence of mitotic cells and general organization of whole LoKI-off ( D ) or LoKI-on ( E ) zebrafish embryos treated with 250 nM CLP-BI2536. EMTB (white and top inset), SNAP (magenta and mid inset), and mitotic cells (cyan and bottom inset) are shown. ( F ) Graph depicting mitotic index measurements for LoKI-expressing embryos; LoKI-off, n = 23, LoKI-on, n = 20, ****p<0.0001. Each point represents % of cells per individual embryo ( n ). Experiments were conducted at least three times (N = 3) and P values were calculated by unpaired two-tailed Student’s t-test. Data are mean ± s.e.m.

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Microinjection, Live Cell Imaging, Expressing, Marker, Two Tailed Test

Journal: eLife

Article Title: Subcellular drug targeting illuminates local kinase action

doi: 10.7554/eLife.52220

Figure Lengend Snippet:

Article Snippet: Peptide, recombinant protein , Plk1, active , SignalChem , P41-10H , .

Techniques: Control, shRNA, Stable Transfection, Selection, Knock-Out, Transfection, Bicinchoninic Acid Protein Assay, DNA Purification, Recombinant, Plasmid Preparation, Modification, Software, Cloning

Journal: Pancreas

Article Title: The Role of p21-Activated Kinases in Pancreatic Cancer

doi: 10.1097/mpa.0000000000000276

Figure Lengend Snippet: FIGURE 3. Possible PAK1 and PAK4 effector functions in pancreatic carcinogenesis. A, The pathways shown play a crucial role in pancreatic cancer, and PAK1 may act as a possible linker or driver. p21-Activated kinase 1 can increase pancreatic cancer metastasis by increasing cytoskeletal dynamics through activation of MLCK, LIMK, and Arpc1b, cell migration through activation of Paxillin and the MAPK/JNK pathway; and EMT through activation of Snail/Slug and the fibronectin/NF-κB pathway. B, p21-Activated kinase1 can increase pancreatic cancer growth by inducing cell cycle transit through activation of Plk1 and Arpc1b, which then activate Aurora kinases, proliferation through activation of MAPK and NF-κB pathways, survival through the inactivation of Bad, and inhibition of PUMA. p21-Activated kinase 1 can inhibit apoptosis through activation of BimL/DLC1. p21-Activated kinase4 can also increase cell proliferation and cell survival through activation of NF-κB pathway and inactivation of Bad, respectively.

Article Snippet: Copyright © 2015 Wolters Kluwer Plk1 is depleted, preventing DNA replication, but increased Plk1 activity allows mitosis to be maintained, resulting in resistance.

Techniques: Activation Assay, Migration, Inhibition