Journal: bioRxiv

Article Title: Eg5 Inhibitor SB743921 Causes p53-Dependent Cell Cycle Arrest, Senescence and Death in Tumor Cells

doi: 10.1101/2025.01.23.634373

Figure Lengend Snippet: Cytotoxicity of SB743921 against mitotic inhibitors in cells with the wild type (WT) and knocked out p53 ( TP53-/- ). A , IC50 values (mean ± SD, n=3) of A549, MCF7 and HCT116 exposed to various mitotic inhibitors for 7 days were obtained based on the SRB assay. B , Proliferative and colony formation ability of A549 WT and A549 TP53-/- after 7 d or 14 d from the removal of SB743921 (SB) or Ixabepilone (Ixa). Cells were exposed to drugs for 72 h and then incubated in fresh media the following 7 d or 14 d. C, Cell cycle distribution of A549 WT and A549 TP53-/- ( left panels ) and MCF7 WT and MCF7 TP53-/- ( right panels ) treated with SB (10 nM) or Ixa (25 nM) for 24 h ( upper panels ) and 72 h ( bottom panels ).

Article Snippet: The following drugs were used: KSP (Eg5) inhibitor SB743921 (SB, Medchem Express), Ixabepilone (Ixa, Swords laboratories), Paclitaxel (Pacl, Fujian South Pharmaceutical), Vincristine (Vinc, Teva), Nocodazole (Noc, Calbiochem), MDM2 inhibitor Nutlin-3 (SelleckChem), Bcl-2/Bcl-xL/Bcl-w inhibitor ABT-263 (ApexBio) and Doxorubicin (SelleckChem).

Techniques: Sulforhodamine B Assay, Incubation

Journal: bioRxiv

Article Title: Eg5 Inhibitor SB743921 Causes p53-Dependent Cell Cycle Arrest, Senescence and Death in Tumor Cells

doi: 10.1101/2025.01.23.634373

Figure Lengend Snippet: p53-mediated cell cycle inhibitory and proapoptotic pathways are activated in response to the exposure of SB743921 (SB) or Ixabepilone (Ixa). A, Immunoblotting analysis of MCF7 and A549 treated with SB or Ixa in different concentrations (SB 0.5 nM and 10 nM, Ixa 2 nM and 25 nM) for 24 h. B, C , qPCR analysis of MCF7 and A549 respectively treated with SB (10 nM for MCF7, 3 nM for A549) or Ixa (25 nM) for 16 h (A549) or 24 h (MCF7).

Article Snippet: The following drugs were used: KSP (Eg5) inhibitor SB743921 (SB, Medchem Express), Ixabepilone (Ixa, Swords laboratories), Paclitaxel (Pacl, Fujian South Pharmaceutical), Vincristine (Vinc, Teva), Nocodazole (Noc, Calbiochem), MDM2 inhibitor Nutlin-3 (SelleckChem), Bcl-2/Bcl-xL/Bcl-w inhibitor ABT-263 (ApexBio) and Doxorubicin (SelleckChem).

Techniques: Western Blot

Journal: bioRxiv

Article Title: Eg5 Inhibitor SB743921 Causes p53-Dependent Cell Cycle Arrest, Senescence and Death in Tumor Cells

doi: 10.1101/2025.01.23.634373

Figure Lengend Snippet: SB743921 induces irreversible proliferative arrest and predominant death of A549 TP53-/- cells. A , B , Cell cycle distribution after 6 days after removal of SB743921 (SB) or Ixabepilone (Ixa). Cells were exposed to the drugs for 24 h, washed and incubated for 3 or 6 d in fresh media. C , Confocal microscopy images of EdU-treated samples after a week since the removal of SB or Ixa. Cells were exposed to drugs for 72 h before the washout. D , Single cell concentrations in flow cytometry analysis as a percentage of the untreated control samples. *, P < 0.05, **, P < 0.01; ns – not significant.

Article Snippet: The following drugs were used: KSP (Eg5) inhibitor SB743921 (SB, Medchem Express), Ixabepilone (Ixa, Swords laboratories), Paclitaxel (Pacl, Fujian South Pharmaceutical), Vincristine (Vinc, Teva), Nocodazole (Noc, Calbiochem), MDM2 inhibitor Nutlin-3 (SelleckChem), Bcl-2/Bcl-xL/Bcl-w inhibitor ABT-263 (ApexBio) and Doxorubicin (SelleckChem).

Techniques: Incubation, Confocal Microscopy, Single Cell, Flow Cytometry, Control

Journal: bioRxiv

Article Title: Eg5 Inhibitor SB743921 Causes p53-Dependent Cell Cycle Arrest, Senescence and Death in Tumor Cells

doi: 10.1101/2025.01.23.634373

Figure Lengend Snippet: SB743921 depletes pro-replicative signaling causing irreversible arrest, while p53 is crucial for G1/S arrest in WT cells. A , Immunoblotting analysis in A549 WT and A549 TP53-/- cells 6 days after removal of SB743921 (SB) or Ixabepilone (Ixa). Cells were exposed to drugs for 24 h, then washed and incubated for 6 d in fresh media. C , qPCR analysis of A549 WT and A549 TP53-/- expression after 6 days since the removal of SB or Ixa. Cells were exposed to drugs for 24 h before the washout.

Article Snippet: The following drugs were used: KSP (Eg5) inhibitor SB743921 (SB, Medchem Express), Ixabepilone (Ixa, Swords laboratories), Paclitaxel (Pacl, Fujian South Pharmaceutical), Vincristine (Vinc, Teva), Nocodazole (Noc, Calbiochem), MDM2 inhibitor Nutlin-3 (SelleckChem), Bcl-2/Bcl-xL/Bcl-w inhibitor ABT-263 (ApexBio) and Doxorubicin (SelleckChem).

Techniques: Western Blot, Incubation, Expressing

Journal: bioRxiv

Article Title: Eg5 Inhibitor SB743921 Causes p53-Dependent Cell Cycle Arrest, Senescence and Death in Tumor Cells

doi: 10.1101/2025.01.23.634373

Figure Lengend Snippet: SB743921 induces irreversible senescence in A549 WT and SB743921-treated senescent cells can be targeted by senolytics, such as ABT-263. A , Light microscopy images of A549 WT 6 days after the removal of SB743921 (SB) or Ixabepilone (Ixa). Cells were fixed and then stained with solution for the SA-β-Gal detection. B, Quantitative analysis of stained A549 WT and A549 TP53-/- cells as demonstrated in A . Data were compared using the ANOVA test; **, P < 0.01, ****, P < 0.0001; ns – not significant. C, Crystal violet staining of A549 WT and A549 TP53-/- treated with SB or Ixa alone or together with 3 uM ABT-263.

Article Snippet: The following drugs were used: KSP (Eg5) inhibitor SB743921 (SB, Medchem Express), Ixabepilone (Ixa, Swords laboratories), Paclitaxel (Pacl, Fujian South Pharmaceutical), Vincristine (Vinc, Teva), Nocodazole (Noc, Calbiochem), MDM2 inhibitor Nutlin-3 (SelleckChem), Bcl-2/Bcl-xL/Bcl-w inhibitor ABT-263 (ApexBio) and Doxorubicin (SelleckChem).

Techniques: Light Microscopy, Staining

Journal: Cancers

Article Title: Inducing Mitotic Catastrophe as a Therapeutic Approach to Improve Outcomes in Ewing Sarcoma.

doi: 10.3390/cancers15204911

Figure Lengend Snippet: Figure 1. An in silico bioinformatics screen identifies mitotic proteins essential for EWS growth. (A) The DepMap portal was used to access the RNA expression data across different cancer cell lines. Expression in Ewing sarcoma is highlighted in red. Capillary-based analysis of protein lysates from EWS cell lines indicating expression of (B) KIF11 and AURKA and (C) KIF15 and TPX2 protein levels. The uncropped blots are shown in Figure S3.

Article Snippet: After this denaturation step, the prepared samples; blocking reagent; 1:50 diluted primary antibodies KIF11 or Eg5 (Cell Signaling Technology, Danvers, MA, USA, Cat# 4203, RRID:AB_10545760), p-KIF11 (Thermo Fisher Scientific Cat# PA5-105186, RRID:AB_2816659), KIF15 (Proteintech, Rosemont, IL, USA, Cat# 55407-1- AP, RRID:AB_11182836), AURKA (Novus Cat# NBP1-51843SS, RRID:AB_11018019), pAURKA (Novus Cat# NBP3-05434) PARP(46D11) (Cell Signaling Technology Cat# 9532, RRID:AB_659884), and β-actin (Cell Signaling Technology Cat# 12262, RRID:AB_2566811); HRP-conjugated secondary antibodies anti-rabbit (Biotechne, Minneapolis, MN, USA, #DM-001) and anti-mouse (Biotechne, #DM-002); and chemiluminescent substrate were dispensed into designated wells in an assay plate.

Techniques: In Silico, RNA Expression, Expressing

Journal: Cancers

Article Title: Inducing Mitotic Catastrophe as a Therapeutic Approach to Improve Outcomes in Ewing Sarcoma.

doi: 10.3390/cancers15204911

Figure Lengend Snippet: Figure 2. Synergistic inhibition of EWS growth by VIC-1911 and different KIF11 inhibitors. Dose– response curves, dose–response matrix, and heat map indicating synergy scores in TC-71 EWS cell

Article Snippet: After this denaturation step, the prepared samples; blocking reagent; 1:50 diluted primary antibodies KIF11 or Eg5 (Cell Signaling Technology, Danvers, MA, USA, Cat# 4203, RRID:AB_10545760), p-KIF11 (Thermo Fisher Scientific Cat# PA5-105186, RRID:AB_2816659), KIF15 (Proteintech, Rosemont, IL, USA, Cat# 55407-1- AP, RRID:AB_11182836), AURKA (Novus Cat# NBP1-51843SS, RRID:AB_11018019), pAURKA (Novus Cat# NBP3-05434) PARP(46D11) (Cell Signaling Technology Cat# 9532, RRID:AB_659884), and β-actin (Cell Signaling Technology Cat# 12262, RRID:AB_2566811); HRP-conjugated secondary antibodies anti-rabbit (Biotechne, Minneapolis, MN, USA, #DM-001) and anti-mouse (Biotechne, #DM-002); and chemiluminescent substrate were dispensed into designated wells in an assay plate.

Techniques: Inhibition

Journal: Cancers

Article Title: Inducing Mitotic Catastrophe as a Therapeutic Approach to Improve Outcomes in Ewing Sarcoma.

doi: 10.3390/cancers15204911

Figure Lengend Snippet: Figure 6. Analysis of protein expression post-drug treatment. (A) CHLA-10 and (B) TC-71 cells treated with drugs were assessed for changes in protein expression 24 h post-treatment via capillary electrophoresis-based Wes analysis. Increased protein levels of KIF11, p-KIF11Thr926 AURKA, and p-AURKAThr288 were observed for the drug combination group, whereas KIF15 levels were noticeably lower. Similarly, enhanced cleaved-PARP expression was observed with the combination treatment. The uncropped blots are shown in Figures S8 and S9.

Article Snippet: After this denaturation step, the prepared samples; blocking reagent; 1:50 diluted primary antibodies KIF11 or Eg5 (Cell Signaling Technology, Danvers, MA, USA, Cat# 4203, RRID:AB_10545760), p-KIF11 (Thermo Fisher Scientific Cat# PA5-105186, RRID:AB_2816659), KIF15 (Proteintech, Rosemont, IL, USA, Cat# 55407-1- AP, RRID:AB_11182836), AURKA (Novus Cat# NBP1-51843SS, RRID:AB_11018019), pAURKA (Novus Cat# NBP3-05434) PARP(46D11) (Cell Signaling Technology Cat# 9532, RRID:AB_659884), and β-actin (Cell Signaling Technology Cat# 12262, RRID:AB_2566811); HRP-conjugated secondary antibodies anti-rabbit (Biotechne, Minneapolis, MN, USA, #DM-001) and anti-mouse (Biotechne, #DM-002); and chemiluminescent substrate were dispensed into designated wells in an assay plate.

Techniques: Expressing, Electrophoresis

Journal: iScience

Article Title: Compartment-specific antibody correlates of protection to SARS-CoV-2 Omicron in macaques

doi: 10.1016/j.isci.2024.110174

Figure Lengend Snippet:

Article Snippet: SARS-CoV-2 EG5 Spike , Sino Biological , 40589-V08H55.

Techniques: Recombinant, Isolation, Cell Isolation, Chromatography, Software, Luminex