aurka inhibitor mln8237 solution  (MedChemExpress)

Journal: bioRxiv

Article Title: Mitotic CDK4/6 activity sustains spindle checkpoint signalling to prevent mitotic slippage and genomic instability

doi: 10.1101/2025.09.13.675969

Figure Lengend Snippet: a, HCT116 cells were synchronised in prometaphase using thymidine-nocodazole treatment, followed by 9 hours of nocodazole incubation. Cells were treated with Palbo or a CDK1-specific inhibitor, RO-3306 (CDK1i, 10 µM), with or without MG132. Samples were collected at 3 and 7 hours after treatment for flow cytometry and immunoblot analysis. Mitotic exit (decrease in pH3-positive cells) induced by Palbo, but not by CDK1i, was effectively blocked by proteasome inhibition. Data are means ± s.d. (N = 3); two-way ANOVA. Western blot analysis shows retention of highly phosphorylated Cdc27 and CycB1 in MG132-treated cells. b, Cells were synchronised in prometaphase as above and treated with Palbo or vehicle control in the presence or absence of the APC/C inhibitor, proTAME (25 µM). Flow cytometry analysis revealed that proTAME blocked mitotic exit induced by CDK4/6 inhibition, as evidenced by retention of pH3-positive cells with high CycB1 levels. c, HCT116 cells were arrested in mitosis using nocodazole and co-treated with MG132 to prevent mitotic exit. BubR1 and centromere marker CENP-C were visualised by immunofluorescence. BubR1 kinetochore localisation decreased over time upon Palbo treatment, while it remained stable in control conditions. Alisertib (1 µM) served as a positive control for SAC inactivation. Scale bar: 10 µm. Pearson correlation coefficients (PCC) quantify BubR1/CENP-C co-localisation; one-way ANOVA. n = 20, 100, 100, 100 (-); 20, 99, 100, 100 (+Palbo); 21, 100, 98, 100 (+Alisertib), at T0, T1, T2, T3 in a single experient. d, CDK4/6 inhibition reduces KNL1 MELT phosphorylation (pMELT) and BubR1 phosphorylation. Left: Immunoblots show decreased pMELT and BubR1 phosphorylation in cells treated with Palbo, Abema (5 µM), or alisertib in the presence of MG132, correlating with SAC inactivation. Right: Immunofluorescence shows partial reduction of kinetochore pMELT signals after Palbo or Abema treatment. Scale bar: 10 µm.

Article Snippet: For kinase inhibition, we employed CDK4/6 inhibitors—palbociclib (MCE, HY-50767), abemaciclib (MCE, HY-16297A), and ribociclib (MCE, HY-15777)—as well as the CDK1-specific inhibitor RO3306 (Selleck, 872573-93-8; MCE, HY-12529), the Aurora A kinase inhibitor alisertib (MCE, HY-10971) and the MEK inhibitor PD0325901 (Selleck S1036).).

Techniques: Incubation, Flow Cytometry, Western Blot, Inhibition, Control, Marker, Immunofluorescence, Positive Control, Phospho-proteomics

Journal: Science Advances

Article Title: Phosphorylation by Aurora kinase A facilitates cortical-cytoplasmic dynamics of Par-3 in asymmetric division of radial glia progenitors

doi: 10.1126/sciadv.adq3858

Figure Lengend Snippet: ( A ) Par-3 consists of three conserved regions (CRs), the oligomerization domain (CR1), the three PDZ domains (CR2), and the aPKC binding domain (CR3). The predicted AurkA phosphorylation sites are located at Ser 227 and Ser 954 (green), which are conserved across vertebrates. ( B ) Experimental schematic of mRNA injection into one-cell or 16/32-cell stage zebrafish embryos. ( C ) Images of embryonic brain morphology at ~28 hpf. Images were taken at 200× on the Zeiss compound microscope. ( D ) Quantification. Phenotyping rubrics are described in fig. S2. ( E ) Time-lapse sequence of images showing Par-3 and Par-3 phospho mutants (GFP only in the heatmap) in mitotic RGPs at the prophase, metaphase, and telophase. ( F ) Quantification. Relative cytoplasmic Par-3 expression was measured at the prophase, metaphase, and telophase using Fiji. One-way ANOVA test with a post hoc comparison to control (Par-3-GFP). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; ns, not significant. For detailed statistics, see table S1. See table S2 for the representative image signal values compared to the median signal values.

Article Snippet: Zebrafish embryos were treated with an AurkA inhibitor (MedChemExpress, MK-5108, HY-13252, CAS no. 1010085-13-8) from 22 to 24 hpf for various durations before imaging (10, 20, 30, 40, 50, 60, and 120 min).

Techniques: Binding Assay, Phospho-proteomics, Injection, Microscopy, Sequencing, Expressing, Comparison, Control

Journal: Science Advances

Article Title: Phosphorylation by Aurora kinase A facilitates cortical-cytoplasmic dynamics of Par-3 in asymmetric division of radial glia progenitors

doi: 10.1126/sciadv.adq3858

Figure Lengend Snippet: ( A ) Experimental schematic: mRNA microinjections into four-cell Tg[b-actin2:H2B-HaloTag] embryos to achieve sparse labeling. Subsequently, embryos were incubated with Halo Tag ligand-Janelia Flour 646 for 2 hours before live imaging. Twenty-four– to 30-hpf embryos are then mounted on a petri dish with a glass cover for live confocal imaging. Confocal image of a 28-hpf zebrafish forebrain (dorsal view) under a 40× objective. The time interval between each volume of z -stacks is 20 s, and the total acquisition time is 30 min. All images shown are the maximum MIP of three confocal z-stacks (1-μm z -step). ( B to D ) In vivo time-lapse imaging of Par-3-GFP and mCherry-AurkA in mitotic RGPs. Arrows indicate AurkA dynamic movements. Par-3-GFP is asymmetrically segregated to the posterior daughter shortly after division, and there is AurkA colocalization toward the posterior side of the cell. ( E ) Quantification. The bar graph illustrates different patterns of Par-3-AurkA colocalization, while Par-3 undergoes asymmetric distribution in RGPs. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; ns, not significant. For detailed statistics, see table S1.

Article Snippet: Zebrafish embryos were treated with an AurkA inhibitor (MedChemExpress, MK-5108, HY-13252, CAS no. 1010085-13-8) from 22 to 24 hpf for various durations before imaging (10, 20, 30, 40, 50, 60, and 120 min).

Techniques: Labeling, Incubation, Imaging, In Vivo

Journal: Science Advances

Article Title: Phosphorylation by Aurora kinase A facilitates cortical-cytoplasmic dynamics of Par-3 in asymmetric division of radial glia progenitors

doi: 10.1126/sciadv.adq3858

Figure Lengend Snippet: ( A ) Experimental schematic. Embryos are injected with mRNA (~200 pg) at the 16/32-cell stage to achieve sparse labeling, followed by Dld antibody uptake at 20 hpf and treatment with 1 μM AurkA inhibitor (MK-5108) for 10 min to 2 hours, and the 24- to 30-hpf zebrafish forebrain region is imaged. A zoomed-in confocal image of a 28-hpf zebrafish forebrain (dorsal view) under a 40× objective. DNA is marked by H2B-RFP (pseudocolored in blue), Par-3-GFP (green), and internalized Dld (magenta). ( B and C ) Time-lapse sequence of images showing the dynamics of internalized Dld and Par-3-GFP in mitotic RGPs. The time of 3 min denotes the telophase when asymmetry indices are calculated. The time interval between each volume of z -stacks is 20 s, and the total acquisition time is 30 min. All images shown are MIP of three confocal z -stacks (1-μm z -step). (B) Control condition: Par-3-GFP time-lapse image showing the dynamics of internalized Dld and Par-3-GFP in mitotic RGPs. (C) Condition: time-lapse sequence of images of 1 μM AurkA Inhibitor for 50 min in mitotic RGPs. Par-3 and Dld asymmetry was disrupted. ( D to G ) Unpaired t test compared to control. (D) Quantification. Percent relative cytoplasmic Par-3 expression was measured at the prophase, metaphase, and telophase using Fiji. (E) Quantification. Percent relative cortical Par-3 expression was measured at the prophase, metaphase, and telophase using Fiji. (F) Asymmetry indices of Par-3-GFP. The dotted lines indicate the threshold of |0.2| for calling asymmetry. (G) Asymmetry indices of internalized Dld (right) in RGPs expressing different forms of Par-3. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; ns, not significant. For detailed statistics, see table S1. See table S2 for the representative image signal values compared to the median signal values.

Article Snippet: Zebrafish embryos were treated with an AurkA inhibitor (MedChemExpress, MK-5108, HY-13252, CAS no. 1010085-13-8) from 22 to 24 hpf for various durations before imaging (10, 20, 30, 40, 50, 60, and 120 min).

Techniques: Injection, Labeling, Sequencing, Control, Expressing