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: 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: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) A petal plot demonstrated 672 overlapping DEGs identified from five RA synovial tissue datasets ( GSE55457 , GSE55235 , GSE2053 , GSE12021 , and GSE1919 ). DEGs were selected with thresholds of |log2FC| ≥ 0.5 and adjusted p-value < 0.05. (B) The mRNA expression levels of KIF11 in rheumatoid arthritis (RA) and normal control (NC) synovial tissues were analyzed in six independent GEO datasets: GSE1919 , GSE12021 , GSE55235 , GSE55457 , GSE77298 , and GSE2053 . Data are presented as mean ± SD. ***P < 0.001, compared with NC controls. (C) MH7A cells were treated with TNF-α (10 ng/mL) and IL-1β (10 ng/mL) for 24 h, and the expression of KIF11 was determined via western blotting. (D) The relative expression of KIF11 was shown in histogram. ***P < 0.01, compared with untreated group.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Expressing, Control, Western Blot

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) The expression of KIF11 was detected by western blotting. (C) Cell viability was measured by MTT assay at 24, 48, and 72 h post-infection with control shRNA (shCtrl) or KIF11 shRNA (shKIF11) lentivirus. n = 3. ***P < 0.001 vs. Ctrl. shRNA group. (D) Edu incorporation assay. Fluorescence imaging results at 24 and 48 hours. E. The proportion of positively stained cells is displayed in a bar graph, ***P < 0.001. F. Colony formation assay. Representative images (E) and quantification (F) of colony formation assay in Ctrl. shRNA- and KIF11 shRNA infected MH7A cells after two-week culture. ***P < 0.001 vs. Ctrl.shRNA group, ###p < 0.001 vs.untreated cells .

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Expressing, Western Blot, MTT Assay, Infection, Control, shRNA, Fluorescence, Imaging, Staining, Colony Assay

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) Transwell migration assay of shCtrl- and shKIF11-infected MH7A cells at 24 h and 48 h. Representative images (40 × magnification) and migrated cell quantification (right). Data are mean ± SD ( n = 3). *P < 0.05, **P < 0.01 vs. CtrlshRNA group. (B) The expression of IL-1β, IL-6, and IL-8 was detected by ELISA assay in cell supernatant. ***P < 0.001. (C) Rescue of KIF11 expression in knockdown cells. MH7A cells with stable KIF11 knockdown (shKIF11) were transfected with either a human KIF11 cDNA overexpression plasmid (KIF11-OE) or the empty control vector (Pcmv3 vector). After 24 hours, KIF11 protein expression was analyzed by Western blotting. β-actin served as a loading control. (D) The relative KIF11 protein levels were shown in histogram. Data are presented as mean ± SD (n = 3). ***P < 0.001 vs. the vector-transfected shKIF11 group. (E) Rescue of pro-inflammatory cytokine secretion. The cell culture supernatants from the experiment described in (D) were collected, and the concentrations of TNF-α, IL-1β, IL-6, and IL-8 were measured by ELISA. Data are presented as mean ± SD (n = 3). ***p < 0.001 vs. the control vector-transfected shKIF11 group.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Transwell Migration Assay, Infection, Expressing, Enzyme-linked Immunosorbent Assay, Knockdown, Transfection, Over Expression, Plasmid Preparation, Control, Western Blot, Cell Culture

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) KIF11 knockdown inhibited NF-κB p65 phosphorylation. MH7A cells were infected with control shRNA (shCtrl) or KIF11 shRNA (shKIF11) lentivirus for 24 hours. Whole-cell lysates were subjected to Western blotting using antibodies against phospho-p65 (p-p65), total p65, and KIF11. β-actin served as a loading control. (B) The histogram shows the relative ratio of p-p65 to total p65. Data are presented as mean ± SD (n = 3). ***P < 0.001 vs. shCtrl group. (C) KIF11 knockdown impaired nuclear translocation of NF-κB p65. Cytoplasmic and nuclear fractions were extracted from shCtrl- and shKIF11-infected MH7A cells. The distribution of p65 in the cytoplasm (Cyto) and nucleus (Nuc) was analyzed by Western blotting. Lamin B1 and α-tubulin were used as markers for nuclear and cytoplasmic fractions, respectively. (D) The relative abundance of p65 in the nuclear and cytoplasmic fractions were shown in histograms, normalized to their respective loading controls. Data are presented as mean ± SD (n = 3). ***P < 0.001 vs. shCtrl group.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Knockdown, Phospho-proteomics, Infection, Control, shRNA, Western Blot, Translocation Assay

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: A. KIF11 was knock down in M1 macrophages for 24 h. B. The histogram of gray value was shown IN Ctrl. shRNA or KIF11 shRNA lentiviruses infected M1 macrophages. **P < 0.01. Interference with KIF11 in M1 macrophages decreased the expression of CD86. The intensity of surface molecules CD14 and CD86 (C) and CD86(D) were determined by flow cytometry in THP-1-derived M0 and M1 cells, shKIF11 lentivirus or ctrl. shRNA lentivirus infected M1 macrophages. E. The histogram of CD86 and CD80 expression was shown. ***P < 0.001.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Knockdown, shRNA, Infection, Expressing, Flow Cytometry, Derivative Assay

Journal: Journal of translational medicine

Article Title: Integrative analysis of T cell-mediated tumor killing-related genes reveals KIF11 as a novel therapeutic target in esophageal squamous cell carcinoma.

doi: 10.1186/s12967-025-06178-y

Figure Lengend Snippet: Fig. 6 Identification of prognostic risk genes. a Correlation analysis among the 8 best prognostic genes in GSE53625. b KM survival analysis displayed high the expression of KIF11correlated with poor prognosis in ESCC patients. c The correlation of KIF11 mRNA expression with tumor cell purity (left) and the degree of CD8+ T cell infiltration (right) in ESCC was analyzed with TIMER 2.0. d mIHC stained by a ESCC tissue microarray. Representative pictures (left) of mIHC assay and the summarized results (right) are shown (n: 70 patients). The data are presented as the means ± standard deviations (SD). Statistical analysis is performed with Pearson correlation analysis (a), Log-rank test (b), Wilcoxon test (c) and Student’s t-test (d). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: Following antibodies were used for Western blotting: KIF11 (A7907, Abclonal, China) and β-actin (Ab8226, Abcam, USA).

Techniques: Expressing, Staining, Microarray

Journal: Journal of translational medicine

Article Title: Integrative analysis of T cell-mediated tumor killing-related genes reveals KIF11 as a novel therapeutic target in esophageal squamous cell carcinoma.

doi: 10.1186/s12967-025-06178-y

Figure Lengend Snippet: Fig. 7 KIF11 increases proliferation and migration of ESCC in vitro. a Western blot performed to assess KIF11 expression in YES2, KYSE30, KYSE150, KYSE410, KYSE450, and KYSE510 cells. (b-c) KIF11 levels in KYSE30 and KYSE450 cells transfected with shRNA (sh1, sh2) and control (NC) were analyzed using Western blot. d–e CCK-8 assays evaluating the proliferation capacity of KIF11-depleted KYSE30 and KYSE450 cells. f Colony formation assays to demonstrate changes in proliferation in KIF11-depleted KYSE30 and KYSE450 cells. g Statistical analysis of colony formation ability in KIF11-depleted KYSE30 and KYSE450 cells. h, i Representative micrographs showed the wound healing efficiency of KYSE30 and KYSE450 cells with reduced KIF11 expression using specific shRNA. j Statistical analysis of the scratch wound healing rate in KIF11-depleted KYSE30 and KYSE450 cells. The data are presented as the means ± standard deviations (SD) of three independent experiments. Statistical analysis is performed with Student’s t-test (d, e, g, j). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: Following antibodies were used for Western blotting: KIF11 (A7907, Abclonal, China) and β-actin (Ab8226, Abcam, USA).

Techniques: Migration, In Vitro, Western Blot, Expressing, Transfection, shRNA, Control, CCK-8 Assay

Journal: Journal of translational medicine

Article Title: Integrative analysis of T cell-mediated tumor killing-related genes reveals KIF11 as a novel therapeutic target in esophageal squamous cell carcinoma.

doi: 10.1186/s12967-025-06178-y

Figure Lengend Snippet: Fig. 8 KIF11 functions as a determinant regulating the growth and metastasis of ESCC cells in vivo. a KIF11-depleted KYSE30 and control KYSE30 cells were subcutaneously injected in BALB/C nude mice. After 4 weeks, the tumors were excised and photographed. b Tumor growth curves were plotted for the mice bearing KYSE30 cells. c Tumor weights were shown for the mice bearing KYSE30 cells. (n = 8). The data are presented as the means ± standard deviations (SD). Statistical analysis is performed with Student’s t-test (c). ***P < 0.001, ****P < 0.0001

Article Snippet: Following antibodies were used for Western blotting: KIF11 (A7907, Abclonal, China) and β-actin (Ab8226, Abcam, USA).

Techniques: In Vivo, Control, Injection