barasertib hqpa  (MedChemExpress)

Journal: bioRxiv

Article Title: Local fluidization of an active cytoplasmic gel partitions large cells

doi: 10.64898/2026.08.11.744254

Figure Lengend Snippet: a , Immunofluorescence images of a Xenopus laevis zygote fixed after the first mitosis, viewed from the animal pole. CPC (anti-INCENP-AF647) localized to the boundary between sister asters (microtubules imaged with anti-tubulin-AF488) and co-localized with an ER-depleted zone (ER labeled with anti-LNPK-AF568). b , Live images of a medaka embryo during 2-cell to 4-cell stage, showing dynamics of microtubules (HiLyte 647-tubulin), CPC (mCherry-Ol-Aurora B) and F-actin (Lifeact-EGFP) from mitosis to cytokinesis. c , CPC (anti-INCENP-AF647) localization in Xenopus egg extract co-imaged with microtubules (EB1-mApple), F-actin (Lifeact-GFP), and mitochondria (MitoView 405) at late anaphase. Top: untreated. Bottom: treated with 0.5 µM barasertib-HQPA. d , Example trajectories of two PEG-coated Fluoresbrite beads of 0.5 µm diameter in particle-tracking microrheology. Scale bar, 5 µm. e , Sampled areas in anaphase cycling egg extract with different cytoskeletal composition. f , Ensemble-averaged pairwise mean-squared displacement (MSD pair ) as a function of time lag τ for particles within each of the three indicated areas in e . g , Apparent diffusion coefficient (D) calculated from MSD pair between adjacent beads. For actin-intact controls, 8 areas (65x65 µm 2 ) in each category were sampled from independent experiments with 3 different egg extracts. The same extracts were used to prepare latrunculin B-added samples, where 20 areas were sampled in microtubule asters and cytoplasm, respectively. Data are mean ± SD. Also see . h , Local D calculated within overlapping 42x42 µm 2 grids, locally averaged, and overlaid onto microtubule images (EB1-mApple). Examples shown are microtubule asters growing against microtubule-free cytoplasm, in actin-intact egg extract (top) or extract treated with 10 µM LatB (bottom). Also see . All scale bars except d , 200 µm.

Article Snippet: Barasertib-HQPA (MedChemExpress, HY-10126) was prepared as an aqueous stock rather than a DMSO stock to avoid perturbing microtubule dynamics by DMSO addition.

Techniques: Immunofluorescence, Labeling, Diffusion-based Assay

Journal: bioRxiv

Article Title: Local fluidization of an active cytoplasmic gel partitions large cells

doi: 10.64898/2026.08.11.744254

Figure Lengend Snippet: a , CPC localization co-imaged with microtubules, F-actin, and mitochondria, from late anaphase to the onset of the following mitosis. b , Mitotic dynamics of actin-intact cycling egg extract treated with 2 μM barasertib-HQPA. Note the lack of spindle assembly or centrosome separation. c , Bottom: timed local addition of barasertib-HQPA into cycling extract reaction with visualization of CPC, microtubules, and F-actin. As the extract reached anaphase, 0.2-0.3 μl of 0.5 mM barasertib-HQPA dissolved in egg extract was pipetted into the reaction adjacent to this pair of asters. Pipetting occurred at approximately 16 min. Top: control reaction without perturbation. d , Similar as c ; bottom: local addition of barasertib-HQPA at approximately 11 min, visualizing mitochondria (MitoView Fix 640), microtubules, and F-actin. e , Dynamics of CPC and mitochondria (MitoView 405) in cycling egg extract where F-actin was depolymerized with 10 μM LatB. All scale bars, 200 μm. Time 0 = first frame with assembled spindles (and fully disassembled nuclei). Imaging probes include EB1-mApple (microtubule +tip), Lifeact-GFP (F-actin), anti-INCENP-AF647 (CPC) and MitoView 405 or MitoView Fix 640 (mitochondria).

Article Snippet: Barasertib-HQPA (MedChemExpress, HY-10126) was prepared as an aqueous stock rather than a DMSO stock to avoid perturbing microtubule dynamics by DMSO addition.

Techniques: Control, Imaging

Journal: Nucleic Acids Research

Article Title: AURKB-driven dissolution of CIZ1–RNA assemblies from the inactive X chromosome in mitosis

doi: 10.1093/nar/gkag018

Figure Lengend Snippet: Dissolution of CIZ1–Xi assemblies in mitosis. ( A ) Model of CIZ1–RNA assemblies surrounding and protecting the modification status of underlying chromatin [ , ]. ( B ) Illustrative immunofluorescence images of female murine D3T3 cells stained for CIZ1 via N-terminal epitopes (CIZ1-N, red, detected with pAb 1794), revealing large protein assemblies at the inactive X chromosome (white arrows) that are not detected in mitosis. DNA is shown in blue, bar is 5 µm. ( C ) Diagram illustrating loss in mitosis and the early G1 phase window during which reformation of CIZ1–Xi assemblies takes place, determined previously using cells that were synchronized in G1 phase by release from arrest with nocodazole . ( D ) Map showing conserved putative AURKB phosphorylation sites between murine and human CIZ1 (circles) displayed on full-length murine CIZ1 ( NP_082688.1 .) The location of epitopes of CIZ1 antibodies used throughout is shown above. Conserved prion-like domains (PLD1 and PLD2) are in red , zinc fingers 1–3 in cyan (ZnF_C2H2 SM00355, ZF_C2H2 sd00020, and ZF_C2H2 sd00020), acidic region (Ac) in yellow, matrin-3 homology domain (MH3) in orange (ZnF_U1, smart00451), and h37/m38 amino-acid C-terminal tail in blue. The sequence context and identity of three conserved AURKB phosphorylation sites in the extreme C-terminus are shown. ( E ) Frequency of cells with CIZ1–Xi assemblies (red) or nucleus-wide SAFA (blue) in cells passing through the stages of mitosis indicated, for D3T3 cells and female primary embryonic fibroblasts (PEFs at p3), in the presence and absence of the AURKB kinase inhibitor barasertib at 0.1 and 1 µM. Results show the average of 3–4 independent replicates within one experiment for each line, with SEM. n indicates the number of nuclei inspected (PEF grey, 3T3 black). Statistical analysis of CIZ1–Xi frequency in anaphase cells shows one-way ANOVA with Tukey post hoc test within each cell type, where * <.05, ** <.01, *** <.001. Below, example immunofluorescence images of D3T3 cells through mitosis, with and without 1 µM barasertib. Cells were stained for the N-terminal domains of CIZ1 (CIZ1-N, red) and SAFA (green). DNA is shown in blue, bar is 5 microns. ( F ) Upper histogram shows the proportion of cells with CIZ1-marked Xi in cycling populations of female D3T3 cells after the indicated times exposed to 300 nM Okadaic acid, visualized via CIZ1-N (red). Lower, histograms show the effect of the indicated concentrations of tautomycin for 15 h, stained for CIZ1-N or the ‘tail’ epitope in the C-terminal end of CIZ1 (CIZ1-C, rabbit pAb). Comparison of technical replicates is by t-test, where * <.05, ** <.01, *** <.001. Error bars show SEM. Below, example images showing H3K27me3-marked Xi chromatin in cells stained for CIZ1-N or CIZ1-C at 15 h with or without tautomycin. Bar is 5 microns.

Article Snippet: Barasertib , Selleck , Cat#AZD1152-HQPA.

Techniques: Dissolution, Modification, Immunofluorescence, Staining, Phospho-proteomics, Zinc-Fingers, Sequencing, Comparison

Journal: Cancer Discovery

Article Title: Epigenetic and Transcriptional Programs Define Osteosarcoma Subtypes and Establish Targetable Vulnerabilities

doi: 10.1158/2159-8290.CD-25-0237

Figure Lengend Snippet: EOD and LOD osteosarcomas have differential drug response. A, IC 50 curves showing the response to trametinib in vitro ( n = 16). The cell lines are colored based on the subtype (EOD/LOD). Drug response was measured after 7 days of treatment. Cell lines are listed in the order they appear, highest to lowest, at the highest concentration. B, Differential drug response to trametinib ( n = 15). Mean with SD with a Mann–Whitney test P value (**, 0.0022). C, IC 50 curves showing the response to AZD1152 in vitro ( n = 16). The cell lines are colored based on the c-MYC protein levels (c-MYC high/c-MYC low). D, Relationship between the c-MYC protein level and the IC 50 of the response to AURKBi (AZD1152). E, Cell injection and treatment schedule of EOD/LOD tumors. F, EOD tumors respond to AURKBi (AZD1152) in vivo in a subcutaneous model. Eight vehicle tumors, nine AZD1152-treated tumors, and nine trametinib-treated tumors. Tumor volume (mm 3 ) measured over time (left). Waterfall plot showing the decrease in tumor volume (center). Analysis of end point (right). Mean with SD with a Mann–Whitney test P value (***, 0.0001. G, LOD tumors respond to trametinib. Ten vehicle tumors, 12 AZD1152-treated tumors, and 12 trametinib-treated tumors. Tumor volume (mm 3 ) measured over time (left). Waterfall plot showing the change in tumor volume (center). Analysis of end point (right). Mean with SD with Wilcoxon test P values = (**, 0.0025; *, 0.0184). H, Strategy to evaluate drug combination in the PDXs. I, PDX (PDX833) shows a better response to combination therapy targeting both epigenetic cell states (AZD1152 and trametinib). Eight vehicle tumors, nine AZD1152-treated tumors, eight trametinib-treated tumors, and eight tumors treated with AZD1152 and trametinib. Tumor volume (mm 3 ) measured over time (left). Waterfall plot showing change in tumor volume (center). End point analysis comparing combination therapy with monotherapy (right). Mean with SD with Mann–Whitney test P values (***, 0.0007 & 0.0002; *, 0.0206). J, PDX (PDX774) shows a better response to combination therapy over time. Mean with SD with Mann–Whitney test P values (**, 0.0047). K, PDX (PDX526) also shows a better response to combination therapy over time. Mean with SD with Mann–Whitney test P values (**, 0.0043). L, PDX (PDX107) also shows a better response to combination therapy over time. Mean with SD with Mann–Whitney test P values (*, 0.0182). See also Supplementary Fig. S8.

Article Snippet: Barasertib-HQPA (AZD2811 or AZD1152, #HY-10126) and trametinib (#HY-10999) were purchased from MedChemExpress.

Techniques: In Vitro, Concentration Assay, MANN-WHITNEY, Injection, In Vivo