Journal: Nature Communications

Article Title: A chemical-genetic system to rapidly inhibit the PP2A-B56 phosphatase reveals a role at metaphase kinetochores

doi: 10.1038/s41467-025-58185-8

Figure Lengend Snippet: A Schematic illustrating kinetochore phosphosites on the BUB complex and KNL1 that are regulated by PP2A-B56. B – C Effects of PP2A-B56 inhibition on the levels of BUB1-pT609, BUB1-pT461 and BUB1 ( B , left) or KNL1-pMELT, KNL1-pRVSF and KNL1 ( B , right) or BUBR1-pT620 and BUBR1 ( C ) at unattached kinetochores, in nocodazole-arrested HeLa-FRT cells with/without directSLiM LIE1 expression and treated with vehicle or rapamycin for 20 min. Kinetochore intensities from 20–30 cells, 2–3 experiments. D Levels of BubR1-pT620 at unattached kinetochores in nocodazole-arrested directSLiM LIE1 cells treated with rapamycin for indicated times. Kinetochore intensities from 30 cells, 3 experiments E Heat map of phosphorylation sites upregulated after PP2A-B56 inhibition in directSLiM LIE1 cells. Each row shows the z-scored intensity of a phosphorylation site ( n = 170). Source data are provided as a Source Data file. Data information: Kinetochore intensities in B – C are normalized to directSLiM LIE1 vehicle condition. Violin plots show the distributions of kinetochore intensities between cells. For each violin plot, each dot represents an individual cell, the horizontal line represents the median and the vertical one the 95% CI of the median, which can be used for statistical comparison of different conditions (see Methods).

Article Snippet: The following primary antibodies (all diluted in 3% BSA in PBS) were used at the final concentration indicated: guinea pig anti-CENP-C (PD030 from Caltag + Medsystems, 1:5000), rabbit anti-BUB1 (A300-373A from Bethyl, 1:1000), mouse anti-BUBR1(8G1) (05-898 from Millipore, 1:1000), rabbit anti-BUBR1 (A300-386A from Bethyl, 1:1000), rabbit anti-KNL1 (ab70537 from abcam, 1:1000), rabbit anti-SKAP(KNSTRN) (HPA042027 from Atlas Antibodies, 1:1000), rabbit anti-HEC1-pSer55 (GTX70017 from Genetex, 1:500), mouse anti-FLAG(M2) (F3165-.2MG from Sigma, 1:1000).

Techniques: Inhibition, Expressing, Comparison

Journal: Nature Communications

Article Title: A chemical-genetic system to rapidly inhibit the PP2A-B56 phosphatase reveals a role at metaphase kinetochores

doi: 10.1038/s41467-025-58185-8

Figure Lengend Snippet: A Table reporting the tested SLiMs with a lower binding affinity for PP2A-B56 in comparison to LIE1. For each peptide sequence, the SLiM is highlighted in bold. B Effects of PP2A-B56 inhibition on the levels of BUBR1-pT620 at unattached kinetochores (top) and on the mitotic cell fate after nuclear envelope breakdown (NEBD, bottom) in HeLa-FRT cells with/without directSLiMs LIE1-7/AAA expression and treated with vehicle or rapamycin. Top graph: Kinetochore intensities from 20 nocodazole-arrested cells, 2 experiments except HeLa-FRT, DMSO: 10 cells. The treatment with vehicle/rapamycin was performed for 20 min prior fixation. Bottom graph: heatmap showing the mean frequencies of cell fate after NEBD in each condition: 2 experiments, 17–50 cells per condition per experiment (see also Supplementary Fig. ). C Table reporting the tested SLiMs with different numbers of acidic residues C-terminal to the LxxIxE motif. The acidic residues restored from LPTI (modified peptide) are highlighted in bold. D Effects of PP2A-B56 inhibition on the levels of BUBR1-pT620 at unattached kinetochores (top) and on the mitotic cell fate after nuclear envelope breakdown (NEBD, bottom) in HeLa-FRT cells with/without directSLiMs LIE1/LIE8-14/AAA expression and treated with vehicle or rapamycin. Top graph: Kinetochore intensities from 20 nocodazole-arrested cells, 2 experiments. The treatment with vehicle/rapamycin was performed for 20 min prior fixation. Bottom graph: heatmap showing the mean frequencies of cell fate after NEBD in each condition: 2 experiments, 42-50 cells per condition per experiment (see also Supplementary Fig. ). E Levels of BubR1-pT620, Bub1-pT609, Knl1-pMELT and Knl1-pRVSF at unattached kinetochores, in nocodazole-arrested HeLa-FRT cells expressing directSLiMs LIE1/LIE9 and treated with vehicle for 20 min. Note that the distributions of directSLiM LIE1 and directSliM LIE9 are also shown in Supplementary Fig. . Kinetochore intensities from 20 to 30 cells, 2–3 experiments. Source data are provided as a Source Data file. Data information: Kinetochore intensities in B, D and E are normalized to directSLiM LIE1 vehicle condition. Violin plots show the distributions of kinetochore intensities between cells. For each violin plot, each dot represents an individual cell, the horizontal line represents the median and the vertical one the 95% CI of the median, which can be used for statistical comparison of different conditions (see Methods).

Article Snippet: The following primary antibodies (all diluted in 3% BSA in PBS) were used at the final concentration indicated: guinea pig anti-CENP-C (PD030 from Caltag + Medsystems, 1:5000), rabbit anti-BUB1 (A300-373A from Bethyl, 1:1000), mouse anti-BUBR1(8G1) (05-898 from Millipore, 1:1000), rabbit anti-BUBR1 (A300-386A from Bethyl, 1:1000), rabbit anti-KNL1 (ab70537 from abcam, 1:1000), rabbit anti-SKAP(KNSTRN) (HPA042027 from Atlas Antibodies, 1:1000), rabbit anti-HEC1-pSer55 (GTX70017 from Genetex, 1:500), mouse anti-FLAG(M2) (F3165-.2MG from Sigma, 1:1000).

Techniques: Binding Assay, Comparison, Sequencing, Inhibition, Expressing, Modification

Journal: Scientific Reports

Article Title: Correlation of BUB1 and BUB1B with the development and prognosis of endometrial cancer

doi: 10.1038/s41598-024-67528-2

Figure Lengend Snippet: Analysis of BUB1 and BUB1B gene differential expression in tumors. ( A ) Analysis of BUB1 gene expression in tumors in TIMER 2.0 database; ( B ) Analysis of BUB1 gene expression analysis in tumors in GEPIA2 database; ( C ) Analysis of BUB1B gene expression analysis in tumors in TIMER 2.0 database; ( D ) Analysis of BUB1B gene expression analysis in tumors in GEPIA2 database. * P < 0.05.

Article Snippet: Samples were incubated with primary antibody (rabbit anti-human BUB1 and BUBIB monoclonal antibody, 1:200) (Sanying Biotechnology Co. Ltd., Wuhan, Hubei, China), followed by incubation with secondary antibody (HRP-labeled goat anti-rabbit II antibody, 1:200) (Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China).

Techniques: Quantitative Proteomics, Gene Expression

Journal: Scientific Reports

Article Title: Correlation of BUB1 and BUB1B with the development and prognosis of endometrial cancer

doi: 10.1038/s41598-024-67528-2

Figure Lengend Snippet: BUB1 and BUB1B expression in EC. ( A ) BUB1 expression in normal endometrium and EC (XianTao database); ( B ) BUB1 expression in normal endometrium and EC (IHC); ( C ) BUB1B expression in normal endometrium and EC (XiaoTao database); ( D ) BUB1B expression in normal endometrium and EC (IHC); ( E ) BUB1 ROC curve in EC; ( F ) BUB1B ROC curve in EC.

Article Snippet: Samples were incubated with primary antibody (rabbit anti-human BUB1 and BUBIB monoclonal antibody, 1:200) (Sanying Biotechnology Co. Ltd., Wuhan, Hubei, China), followed by incubation with secondary antibody (HRP-labeled goat anti-rabbit II antibody, 1:200) (Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Correlation of BUB1 and BUB1B with the development and prognosis of endometrial cancer

doi: 10.1038/s41598-024-67528-2

Figure Lengend Snippet: BUB1 and BUB1B gene variations in EC. ( A ) BUB1 gene alteration in EC; ( B ) BUB1 gene alteration in relation to OS in EC; ( C ) BUB1 gene alteration in relation to PFS in EC. ( D ) BUB1B gene alteration in EC; € BUB1B gene alteration in relation to OS in EC; F. BUB1B gene alteration in relation to PFS in EC.

Article Snippet: Samples were incubated with primary antibody (rabbit anti-human BUB1 and BUBIB monoclonal antibody, 1:200) (Sanying Biotechnology Co. Ltd., Wuhan, Hubei, China), followed by incubation with secondary antibody (HRP-labeled goat anti-rabbit II antibody, 1:200) (Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China).

Techniques:

Journal: Scientific Reports

Article Title: Correlation of BUB1 and BUB1B with the development and prognosis of endometrial cancer

doi: 10.1038/s41598-024-67528-2

Figure Lengend Snippet: BUB1 or BUB1B and EC immune filtration. ( A ) BUB1 expression was associated with TIL abundance in EC; ( B ) BUB1 expression was associated with immunosuppressants in EC; ( C ) BUB1 expression and immunostimulants in EC. ( D ) BUB1B expression was associated with TIL abundance in EC; E. BUB1B expression was associated with immunosuppressants in EC; ( F ) BUB1B expression and immunostimulants in EC.

Article Snippet: Samples were incubated with primary antibody (rabbit anti-human BUB1 and BUBIB monoclonal antibody, 1:200) (Sanying Biotechnology Co. Ltd., Wuhan, Hubei, China), followed by incubation with secondary antibody (HRP-labeled goat anti-rabbit II antibody, 1:200) (Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China).

Techniques: Filtration, Expressing

Journal: Scientific Reports

Article Title: Correlation of BUB1 and BUB1B with the development and prognosis of endometrial cancer

doi: 10.1038/s41598-024-67528-2

Figure Lengend Snippet: The relation between BUB1 and BUB1B expression with clinicopathological characteristics in EC.

Article Snippet: Samples were incubated with primary antibody (rabbit anti-human BUB1 and BUBIB monoclonal antibody, 1:200) (Sanying Biotechnology Co. Ltd., Wuhan, Hubei, China), followed by incubation with secondary antibody (HRP-labeled goat anti-rabbit II antibody, 1:200) (Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Correlation of BUB1 and BUB1B with the development and prognosis of endometrial cancer

doi: 10.1038/s41598-024-67528-2

Figure Lengend Snippet: Influence of BUB1 and BUBIB knockdown on EC cell phenotype. ( A ) The growth of Ishikawa cells in each group; ( B ) The migration rates of Ishikawa cells in each group: blank group (50.99 ± 1.02)%, NC-siRNA group (66.50 ± 7.74)%, BUB1-siRNA group (34.87 ± 4.26)%, and BUB1B-siRNA group (31.96 ± 4.72)%; ( C ) The number of cells that crossed the membrane in each group: blank group (127.3 ± 4.8), NC-siRNA group (125.3 ± 3.9), BUB1-siRNA group (48.6 ± 3.2), and BUB1B-siRNA group (35.0 ± 2.1). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Samples were incubated with primary antibody (rabbit anti-human BUB1 and BUBIB monoclonal antibody, 1:200) (Sanying Biotechnology Co. Ltd., Wuhan, Hubei, China), followed by incubation with secondary antibody (HRP-labeled goat anti-rabbit II antibody, 1:200) (Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China).

Techniques: Knockdown, Migration, Membrane

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: Clinicopathological features of 232 patients with thyroid cancer.

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Expressing, Dissection

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: BUB1 expression with its clinical significance in TC. (A) Association between PFS and BUB1 expression (RNA‐seq) in TC from Kaplan–Meier plotter database. (B) Correlation between BUB1 expression (IHC) and PFS based on a cohort of a total of 232 samples of PTC from tissue microarrays. (C, D) BUB1 expression in normal and TC samples and the relevance with tumour stage in GEPIA database. (E–G) Expression of BUB1 in TC from datasets GSE33630, GSE29265 and GSE65144. (H) BUB1 expression was examined in normal, PTC and ATC tissues through immunohistochemistry staining. (I) BUB1 protein levels among TC cells were detected using WB assays. (* p < 0.05, ** p < 0.01, *** p < 0.001; mean ± standard deviation (SD)).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Expressing, RNA Sequencing, Immunohistochemistry, Staining, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of BUB1 expression with patient prognosis.

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Expressing, Dissection

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: Reduced expression of BUB1 in TC cells suppresses invasive phenotype. (A, B) Verification of BUB1‐siRNA silencing efficacy at mRNA and protein levels. Cell viability assays (C) and colony‐formation assays (D) were measured after BUB1 silencing. (E) The cell cycle of BUB1 silencing in TC cells was determined using flow cytometry. (F–H) Invasion, migration and wound healing assays of BUB1 silence were conducted in TC cells. (* p < 0.05, ** p < 0.01, *** p < 0.001; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Expressing, Flow Cytometry, Migration

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: Nude mice with ATC xenografts exhibit decreased tumorigenesis and slower tumour growth after BUB1 knockdown. The xenografts were developed from BUB1‐WT or BUB1‐KD 8505C cells. (A–E) Body weight, tumour formation rate, tumour weight and tumour volume. (F) H&E staining of tumour tissues. (G) Detection of Ki67 expression performed by immunohistochemistry. (H) The zebrafish xenograft model was employed to evaluate the metastatic ability of 8505C cells, after BUB1 knockdown. (I) Quantification of disseminated tumour foci. (* p < 0.05, ** p < 0.01, *** p < 0.001; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Knockdown, Staining, Expressing, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: The effect of tumour inhibition by BUB1 inhibitor BAY‐1816032 in vitro. (A) We exposed cancer cells to varying BAY‐1816032 concentrations (0, 1.5625, 3.125, 6.25, 12.5, 25 and 50 μM) for 48 h. CCK‐8 assays were employed to investigate cell viability. Colony‐formation assays (B), cell cycle (C), invasion (D), migration (E) and wound healing assays (F) were detected after being treated with BAY‐1816032 in a dose‐dependent manner (0, 2.5, 5 μM). (* p < 0.05, ** p < 0.01, *** p < 0.001; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Inhibition, In Vitro, CCK-8 Assay, Migration

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: The activity of BUB1 inhibition by BAY‐1816032 for tumour growth in ATC in vivo. (A) A BAY‐1816032 oral administration anti‐tumour experiment performed with a mouse xenograft model is shown. We divided the xenografts into the control group and the BAY‐1816032 group after 2 weeks. (B–E) Body weight, tumour weight and tumour volume. (F–G) H&E staining and immunohistochemistry of Ki67 in each group of mice tumour tissues. (H) H&E staining in each group of mice hearts, livers, spleens, lungs and kidneys. (* p < 0.05, ** p < 0.01; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Activity Assay, Inhibition, In Vivo, Control, Staining, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: BUB1 evokes CIN in ATC. (A) A biological process enrichment GO annotation was performed using GSEA grouped by expression of BUB1. (B, C) Gain or loss of the chromosomal segments in 8505C cells after BUB1 knockdown was detected by WES. (D) Immunofluorescent staining of α‐tubulin to detect the formation of abnormal spindle after BUB1 knockdown. (E) The nuclear abnormalities in BUB1‐KD 8505C cells, as revealed by the Giemsa staining, include nucleoplasmic bridges (NPB), micronucleus (MN) and nuclear buds (NBUD). (** p < 0.01, *** p < 0.001; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Expressing, Knockdown, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: BUB1 promotes CIN through interacting with KIF14 in ATC. (A) All significantly enriched gene ontology‐biological process terms from the top 10 enriched genes between high and low BUB1 expression were displayed on a heatmap. KIF14 expression (B–D) and its clinical significance (E) in TC. (F) Correlation analysis of BUB1 and KIF14 expression levels in the GEPIA database. (G) Immunofluorescence assays were conducted to verify the colocalization of BUB1 and KIF14. (H) Lysates derived from Nthy‐ori 3‐1 and 8505C cells were utilized for co‐IP of BUB1 and KIF14. The odds ratio of KIF14/BUB1 protein expression was calculated as KIF 14 IP KIF 14 input ÷ BUB 1 IP BUB 1 input . (I) Verification of KIF14 siRNA silencing efficacy at the mRNA level in 8505C cells. Immunofluorescent staining (J), Giemsa staining (K), and cell cycle distribution analysis (L) were performed after silencing of KIF14. (M) Temporal variation in p‐H3 examined by WB. (* p < 0.05, ** p < 0.01, *** p < 0.001; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Expressing, Immunofluorescence, Derivative Assay, Co-Immunoprecipitation Assay, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: BUB1/KIF14 complex promotes anaplastic thyroid carcinoma progression by inducing chromosome instability

doi: 10.1111/jcmm.18182

Figure Lengend Snippet: BUB1 induces CIN of ATC through phosphorylation of KIF14 at Ser 1292 . (A, B) The effects of BUB1 knockdown and BUB1 inhibitor BAY‐1816032 on KIF14 phosphorylation were detected using Phos‐tag assays. SDS‐PAGE gels were used to conduct WB for total KIF14. The expression level of p‐KIF14 protein was determined by its ratio to the total expression level of KIF14 protein. (C) The potential phosphorylation sites of KIF14 were retrieved from the UniProt website and mutated to alanine (Ala), respectively. (D) The effects of Flag‐tagged KIF14 mutant plasmids on its phosphorylation were conducted by a Phos‐tag assay in 8505C cells. (E) Cell viability assays were measured after transfecting with Flag‐tagged KIF14 mutant plasmids. (F) Lysates derived from 293T cells were utilized for co‐IP of HA‐BUB1 and FLAG‐KIF14. The cells were all transfected with HA‐BUB1 but were transfected with KIF14 ΔSer1292 mutant or WT plasmids, respectively. The odds ratio of FLAG‐KIF14/HA‐BUB1 protein expression was calculated as FLAG − KIF 14 IP FLAG − KIF 14 input ÷ HA − BUB 1 IP HA − BUB 1 input . (G) Lysates derived from 293T cells were utilized for co‐IP of HA‐BUB1 and FLAG‐KIF14. The cells were first transfected with HA‐BUB1 and FLAG‐KIF14 WT plasmids and then treated with BAY‐1816032 in a dose‐dependent manner. The odds ratio of FLAG‐KIF14/HA‐BUB1 protein expression was calculated as FLAG − KIF 14 IP FLAG − KIF 14 input ÷ HA − BUB 1 IP HA − BUB 1 input . (H) The in vitro kinase assays. BUB1 kinase was mixed with purified recombinant KIF14 or KIF14 ΔSer1292 protein, and KIF14 phosphorylation was examined by Phos‐tag assays. The expression level of p‐KIF14 protein was determined by its ratio to the total expression level of KIF14 protein. Immunofluorescent staining (I), Giemsa staining (J) and cell cycle distribution analysis (K) were performed after overexpression of KIF14 ΔSer1292 mutant (* p < 0.05, ** p < 0.01, *** p < 0.001; mean ± SD).

Article Snippet: For WB, the primary antibodies included: BUB1 (A18053, ABclonal, Wuhan, China), KIF14 (26000‐1‐AP, Proteintech), p‐H3 (53,348, CST, USA), Flag (AE005, ABclonal), HA (AE105, ABclonal), GAPDH (10494‐1‐AP, Proteintech).

Techniques: Phospho-proteomics, Knockdown, SDS Page, Expressing, Mutagenesis, Derivative Assay, Co-Immunoprecipitation Assay, Transfection, In Vitro, Purification, Recombinant, Staining, Over Expression