Journal: Prostate cancer and prostatic diseases

Article Title: Discovery of a new candidate drug to overcome cabazitaxel-resistant gene signature in castration-resistant prostate cancer by in silico screening.

doi: 10.1038/s41391-021-00426-0

Figure Lengend Snippet: Fig. 2 AURKB and KIF20A as therapeutic targets of pimozide (PZD). A Venn diagram of genes upregulated in CBZ-resistant DU145CR and downregulated by PZD. B DAVID functional annotation clustering revealed genes related to cell division, chromatin cohesion, and kinetochore were downregulated by PZD. C Kaplan–Meier curves of recurrence-free survival in The Cancer Genome Atlas prostate cancer cohort by high or low AURKB expression. D Kaplan–Meier curves of recurrence-free survival according to KIF20A expression in The Cancer Genome Atlas prostate cancer cohort.

Article Snippet: Human overexpression lysates of AURKB and KIF20A were obtained from OriGene Technologies, Rockville, MD, USA.

Techniques: Biomarker Discovery, Functional Assay, Expressing

Journal: Prostate cancer and prostatic diseases

Article Title: Discovery of a new candidate drug to overcome cabazitaxel-resistant gene signature in castration-resistant prostate cancer by in silico screening.

doi: 10.1038/s41391-021-00426-0

Figure Lengend Snippet: Fig. 3 AURKB / KIF20A expression in cabazitaxel-resistant prostate cancer cells. A AURKB and KIF20A mRNA expression in CBZ-sensitive DU145 and in DU145CR cells. B AURKB or KIF20A protein expression in LNCaP, DU145, DU145CR cells and overexpression lysates of AURKB or KIF20A. C Flow cytometry indicating the percentage of AURKB-positive cells is increased in DU145CR vs. DU145 cells. D KIF20A-positive cells are also increased in DU145CR cells.

Article Snippet: Human overexpression lysates of AURKB and KIF20A were obtained from OriGene Technologies, Rockville, MD, USA.

Techniques: Expressing, Over Expression, Flow Cytometry

Journal: Cell Death & Disease

Article Title: F-box protein Fbxl18 mediates polyubiquitylation and proteasomal degradation of the pro-apoptotic SCF subunit Fbxl7

doi: 10.1038/cddis.2014.585

Figure Lengend Snippet: Ectopically expressed Fbxl7 induces apoptosis. ( a , b ) Hela cells were transfected with either 2- μ g control vector, 1- or 2 -μ g Fbxl7 -V5 plasmid as indicated. After 20 h, 40 h incubation, cells were collected and stained with FITC-Annexin V and propidium iodine prior to analysis by flow cytometry. Data was quantitated and graphed ( b ). * P <0.001 versus vector

Article Snippet: HA mouse antibody, ubiquitin, Cul1, Skp1, and Aurora A antibodies, stress and apoptosis signaling antibody array kits were from Cell Signaling (Danvers, MA, USA).

Techniques: Transfection, Control, Plasmid Preparation, Incubation, Staining, Flow Cytometry

Journal: Virology

Article Title: A single immunization with a recombinant canine adenovirus expressing the rabies virus G protein confers protective immunity against rabies in mice.

doi: 10.1016/j.virol.2006.07.037

Figure Lengend Snippet: Fig. 6. Cell surface expression of RV G on MDCK cells. Cells were either mock- infected (solid histogram) or infected with CAV2 (black histogram), CAV2-RV G (red histogram), or V-RG (green histogram) at a m.o.i. of 1 for 48 h and incubated with recombinant human anti-RV G monoclonal antibody followed by FITC-conjugated anti-human IgG. Surface expression was determined by flow cytometry. Cells infected with CAV2-RV G showed RV G expression levels comparable to those seen in V-RG-infected cells.

Article Snippet: After 20 min, cells were washed twice with PBS containing 10 mM glycine and 1% bovine serum albumin and incubated with recombinant human anti-RVG monoclonal antibody SO57 (1:4000) followed by a FITC-conjugated affinity-purified goat anti-rabbit antibody (1:500; MP Biomedical, Aurora, OH).

Techniques: Expressing, Infection, Incubation, Recombinant, Flow Cytometry

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 1 The protective efficacy of the Ad-AURKA/CDK7 vaccine on various tumors in the preventive model. (A) Schematic diagram of subcutaneous tumors inoculation (Renca, RM-1, MC38 or Hepa1-6) after immunization with Ad-Ctrl or Ad-AURKA/ CDK7 vaccine (n=5 mice per group). (B, F, J and N) Average tumor volumes of each group in the Renca, RM-1, MC38, or Hepa1-6 subcutaneous tumors were measured twice a week. The tumor volume was statistically analyzed 35 days after tumor inoculation. (C, G, K and O) The tumor volume of an individual mouse in each group of Renca, RM-1, MC38, or Hepa1-6 subcutaneous tumors was plotted. (D, H, L and P) Tumor weights were measured at the end of the experiment. (E, I, M and Q) Tumor inhibition rate in the Renca, RM-1, MC38, or Hepa1-6 subcutaneous tumors was, respectively, calculated by (D, H, L and P). The two-tailed independent Student’s t-test was used to analyze two-group comparisons. The data showed as means±SD. The statistical significance levels were set as *p<0.05, **p<0.01 and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; i.m, intramuscular; s.c, subcutaneous.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: Inhibition, Two Tailed Test

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 2 Antitumor efficacy of Ad-AURKA/CDK7 immunization in the Renca subcutaneous tumor model. (A) Schematic diagram showed the overall design of Renca therapeutic subcutaneous tumor (n=5 mice per group). (B) Tumor growth volume of each group after Renca tumor inoculation was measured twice a week. The tumor volume was statistically analyzed 35 days after tumor inoculation. (C) The final tumor volume of different treatment groups on day 35 after Renca tumor implantation. (D) Tumor weights were measured at the end of the experiment. (E) The tumor inhibition rate in (D) is shown. (F, G) The percentages of CD3+ T cells, CD4+ T cells, CD8+ T cells, DCs, NK, Mφ, MDSC, or regulatory T cells (Treg) were from spleens and tumors in each group. One-way analysis of variance was used for comparisons among multiple groups. Data are means±SD. *p<0.05, **p<0.01, ***p<0.001, ****p<0.001 and not significant (ns). Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin- dependent kinase 7; DCs, dendritic cells; i.m, intramuscular; MDSC, myeloid-derived suppressor cells; NK, natural killer; s.c, subcutaneous; TIL, tumor-infiltrating leukocyte.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: Tumor Implantation, Inhibition, Derivative Assay

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 3 The maturation and differentiation of DC subgroups induced by Ad-AURKA/CDK7 vaccine in vivo. Different DC subgroups in spleens were analyzed by flow cytometry at the end of the experiments. (A) The percentages of CD8+CD11c+ DC subsets in immunized mice of each group (n=5), a typical flow cytometry data is displayed from each group. (B, C) Statistical analysis of the ratio of CD11c+ DCs or CD8+CD11c+ DCs. (D) The representative flow cytometry data of the percentage of CD80+CD11c+, CD86+CD11c+, MHC-II+CD11c+, or CD40+CD11c+ DC subsets in spleens of each group. (E–H) Statistical analysis of different DC subgroups in (D). One-way analysis of variance was used for comparisons among multiple groups. Data are means±SD. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin- dependent kinase 7; DCs, dendritic cells.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: In Vivo, Flow Cytometry

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 4 Ad-AURKA/CDK7 treatment induced memory CD8+ T-cell immune response. (A, B) The proliferation capability of antigen-specific CD8+ T cells from mice spleens in each group was detected by the EdU assay after the continuous stimulation with AURKA/CDK7 antigens. (C, D) The number of IFN-γ-secreting T lymphocytes was observed using the ELISpot assay. (E, F) The representative flow cytometry data of CD8+ T cells secreting IFN-γ, TNF-α, or IL-2 in each group after antigen stimulation. (G, H) The co-culture experiment was detected to assess CTL-specific killing ability. (I, J) The proportions of effector memory T cells or central memory T cells in spleens were measured, and typical flow cytometry data was selected from each group. (K) The tumor volume of a single mouse in the Renca subcutaneous tumors was measured twice a week after tumor rechallenge (n=5 mice per group). (L) The survival curve of the Ad-AURKA/CDK7 or Ad-Ctrl group was observed after re-implantation of the Renca tumor (n=10 mice per group). One-way analysis of variance was used for comparisons among multiple groups. Survival analysis was performed using the log-rank (Mantel-Cox) test. Data are means±SD. **p<0.01, ***p<0.001, ***p<0.001 and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; CTL, cytotoxic T lymphocytes; ELISpot, enzyme-linked immunosorbent spot; IFN, interferon; IL, interleukin; TNF, tumor necrosis factor.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: EdU Assay, Enzyme-linked Immunospot, Flow Cytometry, Co-Culture Assay, ELISpot Assay

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 5 Multifunctional CD8+ T cells exerted an indispensable role in the antitumor effects of Ad-AURKA/CDK7 vaccine. (A–D) The percentages of multifunctional CD8+ T cells secreting TNF-α+IFN-γ+, TNF-α+IL-2+, IFN-γ+IL-2+, or TNF-α+IFN-γ+IL-2+ in spleens of each group were detected by flow cytometry after continuous stimulation with AURKA/CDK7 antigens for 96 hours. (E–H) The proportions of tumor-infiltrating multifunctional CD8+ T lymphocytes secreting TNF-α+IFN-γ+, TNF-α+IL-2+, IFN- γ+IL-2+ or TNF-α+IFN-γ+IL-2+ in tumor tissues of each group were analyzed on day 35 after Renca tumor inoculation. (I) The tumor weights of mice in each group were measured at the end of CD8 depletion. (J) Tumor inhibition rates in (I). (K, L) The percentages of CD8+ T cells or CD8+CD11c+ DCs were detected in spleens and tumor tissues of each group in the CD8+ T-cell depletion assay. The two-tailed independent Student’s t-test was used to analyze two-group comparisons. One-way analysis of variance was used for comparisons among multiple groups. The data are shown as means±SD, with n=5 mice per group. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; IFN, interferon; IL, interleukin; TIL, tumor-infiltrating leukocytes; TNF, tumor necrosis factor.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: Flow Cytometry, Inhibition, Depletion Assay, Two Tailed Test

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 6 Ad-AURKA/CDK7 treatment suppressed tumor metastases by activating multifunctional CD8+ T cells. (A) A schematic diagram displayed the design and treatment of lung metastasis. (B) The representative image of lung metastasis nodules in each group. (C) The survival curve of mice in different groups after Renca tumor lung metastasis (n=10 mice per group). (D) The number of metastatic nodules was counted on the lung surface of immunized mice (n=5 mice per group). (E) Typical immunohistochemistry image of the lung-infiltrating CD8+ T cells in each group immunized with different vaccines. The scale was 200 µm. (F, G) The proportion of CD8+ T cells and CD8+CD11c+ DCs in the lungs of treated mice in each group. (H, I) Percentages of multifunctional CD8+ T lymphocytes producing IFN-γ, TNF-α, or IL-2 in spleens and tumor tissues of different groups. (J) The EdU assay was used to detect the proliferation of CD8+ T cells. (K) The number of IFN-γ-secreting T lymphocytes was counted using ELISpot assay. (L) The percentages of tumor-specific killing capability of CTL in each group. One-way analysis of variance was used for comparisons among multiple groups. Survival analysis was performed using the log-rank (Mantel-Cox) test. The data expressed as means±SD. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; CTL, cytotoxic T lymphocyte; DC, dendritic cell; ELISpot, enzyme- linked immunosorbent spot; IFN, interferon; IL, interleukin; i.m, intramuscular; i.v, intravenous; TNF, tumor necrosis factor.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: Immunohistochemistry, Vaccines, EdU Assay, Enzyme-linked Immunospot, ELISpot Assay

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 7 Antitumor efficacy of Ad-AURKA/CDK7 vaccine requires for multi-functional CD8+ T cells in the Renca orthotopic model. (A) Schematic diagram illustrating the design of the Renca orthotopic model and vaccination. (B) Typical image of renal orthotopic tumors in the different vaccine-treated groups at the endpoint of the experiment (n=5 mice per group). (C) The survival curve of each group in the orthotopic model (n=10 mice per group). (D) Tumor weights were calculated by the formula: Tumor mass (g)=left kidney mass − right kidney mass. (E) Tumor inhibition rate in (D). (F) Tumor-infiltrating CD8+ T lymphocytes were captured by immunohistochemical staining in the left kidney and the representative images were shown. The scale was 200 µm. (G, H) Statistical analysis of the proportion of CD8+ T cells or CD8+CD11c+ DCs in renal tumor tissues of each group. (I, J) The percentages of multifunctional CD8+ T lymphocytes secreting IFN-γ+, TNF-α+, IL-2+, TNF-α+IFN-γ+, TNF-α+IL-2+, IFN- γ+IL-2+, or TNF-α+IFN-γ+IL-2+ were detected by flow cytometry in spleens and tumors in each group. (K) The antigen-stimulated proliferation of CD8+ T cells in different groups. (L) The tumor-specific killing capability of CTL was detected by the co-culture experiment. One-way analysis of variance was used for comparisons among multiple groups. Survival analysis was performed using the log-rank (Mantel-Cox) test. The data are shown as means±SD. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; CTL, cytotoxic T lymphocyte; DC, dendritic cell; IFN, interferon; IL, interleukin; i.m, intramuscular; s.c, subcutaneous; TNF, tumor necrosis factor.

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: Functional Assay, Inhibition, Immunohistochemical staining, Staining, Flow Cytometry, Co-Culture Assay

Journal: Journal for immunotherapy of cancer

Article Title: Adenovirus vaccine targeting kinases induces potent antitumor immunity in solid tumors.

doi: 10.1136/jitc-2024-009869

Figure Lengend Snippet: Figure 8 The therapeutic effect induced by Ad-AURKA/CDK7 vaccine in the humanized mice model. (A) Schematic diagram explaining the establishment of the humanized mice model and experiment design. (B) Final tumor volumes were measured in each group on day 35 after OSRC-2 tumor inoculation. (C) Tumor weights of each group at the end of the experiment. (D) Tumor inhibition rate in (C). (E) The representative flow cytometry data of CD8+CD11c+, CD103+CD11c+, CD80+CD11c+, CD86+CD11c+, and HLA-A2+CD11c+ DC subgroups in spleens of each group. (F–J) Statistical analysis of DC subsets in (E) in the spleens of humanized mice. (K–O) The proportions of tumor-infiltrating DC subsets in different groups were detected. (P) The typical flow cytometry image of multi-functional CD8+ T lymphocytes secreting IFN-γ+, TNF-α+, or IL-2+ in spleens in the Ad-Ctrl or Ad-hAURKA/CDK7 treatment group. (Q, S) The percentages of multifunctional CD8+ T cells in (P) in spleens. (R, T) Statistical analysis of the proportion of IFN-γ+CD8+, TNF-α+CD8+, IL-2+CD8+, TNF-α+IFN-γ+CD8+, TNF-α+IL-2+CD8+, IFN-γ+IL-2+CD8+, or TNF-α+IFN-γ+IL-2+CD8+ in antigen-specific CD8+ T lymphocytes of tumor tissues per group. The two-tailed independent Student’s t-test was used to analyze two-group comparisons. One-way analysis of variance was used for comparisons among multiple groups. The data are shown as means±SD, with n=5 mice per group. **p<0.01, ***p<0.001, and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; DC, dendritic cell; IFN, interferon; IL, interleukin; i.m, intramuscular; TIL, tumor-infiltrating leukocytes; TNF, tumor necrosis factor..

Article Snippet: After blocking with 5% skim milk at room temperature for 1 hour, the membranes were incubated overnight at 4°C with primary antibodies targeting AURKA (Proteintech, Cat# 66 757–1- Ig) or CDK7 (Proteintech, Cat# 27 027–1- AP).

Techniques: Inhibition, Flow Cytometry, Functional Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Fc-modification of anti-PcrV gene-encoded antibodies modulates complement-mediated killing of Pseudomonas aeruginosa

doi: 10.3389/fimmu.2025.1618297

Figure Lengend Snippet: Fc-engagement with complement proteins and assessment of DMAb functionality. (A) V2L2-MD DMAb variant binding to C1q (50 µg/mL) coated ELISA plates. Purified V2L2-MD DMAbs were prepared at a concentration of 0.5 mg/mL (diluted 2-fold; group mean ± SD). (B) Purified V2L2-MD DMAb Variant Fc-binding guinea pig complement and probed with FITC-conjugated anti-guinea pig C3 antibody compared to Fc-abrogated TM V2L2, Flu DMAb 2-12C control, and IgG1 Isotype Control (n=2; group mean ± SD). (C) in vivo produced V2L2-MD DMAb C3 deposition was measured utilizing guinea pig complement and probed with a FITC-conjugated anti-guinea pig C3 antibody. All variants were compared to sera from naïve mice (n=5/group; group mean ± SD). Outliers identified (>20,000 MFI, blue) in GraphPad Prism 10 (Q = 1%). Statistical analysis performed on outlier excluded data. (D) PAO1 and (E) PA14 complement-mediated antibody-dependent cellular phagocytic (cADCP) killing assay utilizing purified V2L2 DMAb variants at 200 µg dose. Percent (%) bacterial killing was calculated as ((mean of control - # DMAB variant colonies)/mean control)*100 (n=3/group). Negative percent bacterial killing represents bacterial growth. Statistical analysis performed in GraphPad Prism 10 by one-way ANOVA (* p<0.05 , ** p<0.01 , *** p<0.001 , **** p<0.0001 ).

Article Snippet: Beads were washed with 1X PBS +15mM EDTA for three times and stained with FITC-goat anti-Guinea Pig C3 antibody (MPBio; Cat# 0855385C3 deposition on beads was analyzed by a Cytek Aurora Spectral Flow Cytometer.

Techniques: Variant Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Purification, Concentration Assay, Control, In Vivo, Produced

Journal: International journal of oncology

Article Title: Silencing Aurora-A with siRNA inhibits cell proliferation in human lung adenocarcinoma cells.

doi: 10.3892/ijo.2016.3605

Figure Lengend Snippet: Figure 1. Analysis for expression of AURKA. (A) Expressions of AURKA protein in normal and lung adenocarcinoma tissue. (a) Lung adenocarcinoma (x400); (b) normal tissue (x400). (B) Relationship between AURKA expression and overall survival in lung adenocarcinoma by Kaplan-Meier analysis; (C) RT-PCR analysis for AURKA mRNA levels in NSCLC cells.

Article Snippet: Mouse anti-human AURKA polyclonal antibody was purchased from ProteinTech Group (Wuhan, Hubei, China); mouse anti-human AURKA monoclonal antibody was purchased from Abcam (Abcam, Cambridge, UK); Taq DNA polymerase was obtained from Fermentas, Inc. (Waltham, MA, USA); Lipofectamine 2000, Opti-MEM and the SuperScript III reverse transcriptase (RT) kit were purchased from Invitrogen (Carlsbad, CA, USA); Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and bovine serum albumin were purchased from Sigma Chemical Co. (St. Louis, MO, USA); VCR were purchased from Shenzhen Wan Le Pharmaceutical Co., Ltd. (Shenzhen, Guangdong, China); cell culture media, fetal bovine serum (FBS) and other supplementary materials were purchased from Gibco Co. (Grand Island, NY, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: International journal of oncology

Article Title: Silencing Aurora-A with siRNA inhibits cell proliferation in human lung adenocarcinoma cells.

doi: 10.3892/ijo.2016.3605

Figure Lengend Snippet: Figure 2. Lentivirus infection in lung adenocarcinoma cells. (A) Green fluorescence indicates the successful delivery of siRNA into H1299 and A549 cells, respectively. (B) The AURKA gene silencing in cells. AURKA mRNA expression levels of AURKA-silenced and -non-silenced cells measured by real-time PCR. (C) AURKA protein expression levels of AURKA-silenced and nonsilenced H1299 and A549 cells analyzed by western blotting. **p<0.01, in comparison with scr-siRNA group.

Article Snippet: Mouse anti-human AURKA polyclonal antibody was purchased from ProteinTech Group (Wuhan, Hubei, China); mouse anti-human AURKA monoclonal antibody was purchased from Abcam (Abcam, Cambridge, UK); Taq DNA polymerase was obtained from Fermentas, Inc. (Waltham, MA, USA); Lipofectamine 2000, Opti-MEM and the SuperScript III reverse transcriptase (RT) kit were purchased from Invitrogen (Carlsbad, CA, USA); Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and bovine serum albumin were purchased from Sigma Chemical Co. (St. Louis, MO, USA); VCR were purchased from Shenzhen Wan Le Pharmaceutical Co., Ltd. (Shenzhen, Guangdong, China); cell culture media, fetal bovine serum (FBS) and other supplementary materials were purchased from Gibco Co. (Grand Island, NY, USA).

Techniques: Infection, Fluorescence, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Comparison

Journal: International journal of oncology

Article Title: Silencing Aurora-A with siRNA inhibits cell proliferation in human lung adenocarcinoma cells.

doi: 10.3892/ijo.2016.3605

Figure Lengend Snippet: Figure 3. Effect of AURKA silencing on the proliferation of lung adenocarcinoma cells. (A) Cellomics arrayscan assay. AURKA-siRNA depressed the growth curves of H1299 cells as compared with the scr-siRNA as determined using a Cellomics arrayscan with time-dependent relation. (B) Colony formation assay. Colony formation assay showed that AURKA siRNA decreased the clone numbers of H1299 cells as compared with the scr-siRNA group. (C) MTT assay. The proliferation levels were measured by MTT assay in AURKA-silenced and -non-silenced A549 cells. AURKA-siRNA depressed the growth curves of H1299 cells as compared with the scr-siRNA group. **p<0.01, in comparison with the scr-siRNA group.

Article Snippet: Mouse anti-human AURKA polyclonal antibody was purchased from ProteinTech Group (Wuhan, Hubei, China); mouse anti-human AURKA monoclonal antibody was purchased from Abcam (Abcam, Cambridge, UK); Taq DNA polymerase was obtained from Fermentas, Inc. (Waltham, MA, USA); Lipofectamine 2000, Opti-MEM and the SuperScript III reverse transcriptase (RT) kit were purchased from Invitrogen (Carlsbad, CA, USA); Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and bovine serum albumin were purchased from Sigma Chemical Co. (St. Louis, MO, USA); VCR were purchased from Shenzhen Wan Le Pharmaceutical Co., Ltd. (Shenzhen, Guangdong, China); cell culture media, fetal bovine serum (FBS) and other supplementary materials were purchased from Gibco Co. (Grand Island, NY, USA).

Techniques: Colony Assay, MTT Assay, Comparison

Journal: International journal of oncology

Article Title: Silencing Aurora-A with siRNA inhibits cell proliferation in human lung adenocarcinoma cells.

doi: 10.3892/ijo.2016.3605

Figure Lengend Snippet: Figure 4. The effects of AURKA siRNA on the cell cycle distribution of lung adenocarcinoma cells as determined by flow cytometric analysis. (A) The flow cytometric histogram and the number of cells under each phase of the cell cycle in AURKA-silenced and -non-silenced H1299 and A549 cells. (B) The numerical representation of the cells under each phase of the cell cycle in AURKA-silenced and -non-silenced H1299 and A549 cells.

Article Snippet: Mouse anti-human AURKA polyclonal antibody was purchased from ProteinTech Group (Wuhan, Hubei, China); mouse anti-human AURKA monoclonal antibody was purchased from Abcam (Abcam, Cambridge, UK); Taq DNA polymerase was obtained from Fermentas, Inc. (Waltham, MA, USA); Lipofectamine 2000, Opti-MEM and the SuperScript III reverse transcriptase (RT) kit were purchased from Invitrogen (Carlsbad, CA, USA); Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and bovine serum albumin were purchased from Sigma Chemical Co. (St. Louis, MO, USA); VCR were purchased from Shenzhen Wan Le Pharmaceutical Co., Ltd. (Shenzhen, Guangdong, China); cell culture media, fetal bovine serum (FBS) and other supplementary materials were purchased from Gibco Co. (Grand Island, NY, USA).

Techniques:

Journal: International journal of oncology

Article Title: Silencing Aurora-A with siRNA inhibits cell proliferation in human lung adenocarcinoma cells.

doi: 10.3892/ijo.2016.3605

Figure Lengend Snippet: Figure 5. The effects of AURKA siRNA on the cell apoptosis of lung adenocarcinoma cells as determined by flow cytometric analysis. (A) Flow cytometry analysis shows that knockdown of AURKA expression via AURKA siRNA increased cell apoptosis in H1299 cells. (B) Flow cytometry analysis showed that knockdown of AURKA expression via AURKA siRNA increased cell apoptosis in A549 cells. **p<0.01, in comparison with scr-diRNA group.

Article Snippet: Mouse anti-human AURKA polyclonal antibody was purchased from ProteinTech Group (Wuhan, Hubei, China); mouse anti-human AURKA monoclonal antibody was purchased from Abcam (Abcam, Cambridge, UK); Taq DNA polymerase was obtained from Fermentas, Inc. (Waltham, MA, USA); Lipofectamine 2000, Opti-MEM and the SuperScript III reverse transcriptase (RT) kit were purchased from Invitrogen (Carlsbad, CA, USA); Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and bovine serum albumin were purchased from Sigma Chemical Co. (St. Louis, MO, USA); VCR were purchased from Shenzhen Wan Le Pharmaceutical Co., Ltd. (Shenzhen, Guangdong, China); cell culture media, fetal bovine serum (FBS) and other supplementary materials were purchased from Gibco Co. (Grand Island, NY, USA).

Techniques: Flow Cytometry, Knockdown, Expressing, Comparison

Journal: International journal of oncology

Article Title: Silencing Aurora-A with siRNA inhibits cell proliferation in human lung adenocarcinoma cells.

doi: 10.3892/ijo.2016.3605

Figure Lengend Snippet: Figure 6. Western blotting, BrdU incorporation and flow cytometry analysis. (A) AURKA silencing downregulated RAF-1, CCND2, CCND3, CDK4, PAK4, EGFR and upregulated WEE1 expression in H1299 cells. (B) The DNA synthesis rate was analyzed by BrdU incorporation assay on the 1st and 4th days. Downregulated AURKA in human lung cancer A549 cells displayed changes in DNA synthesis. Silencing AURKA gene expression by siRNA can signifi- cantly enhance chemotherapy sensitivity effect by combining VCR in lung cancer A549 cells. (C) Apoptosis sensitivity of A549 cells towards VCR after downregulation of AURKA. The rate of apoptosis of A549 cells was significantly higher in A549 cells were treated with VCR after transfection with AURKA- siRNA than those treated with VCR after transfection with scr-siRNA. **p<0.01, in comparison with control.

Article Snippet: Mouse anti-human AURKA polyclonal antibody was purchased from ProteinTech Group (Wuhan, Hubei, China); mouse anti-human AURKA monoclonal antibody was purchased from Abcam (Abcam, Cambridge, UK); Taq DNA polymerase was obtained from Fermentas, Inc. (Waltham, MA, USA); Lipofectamine 2000, Opti-MEM and the SuperScript III reverse transcriptase (RT) kit were purchased from Invitrogen (Carlsbad, CA, USA); Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and bovine serum albumin were purchased from Sigma Chemical Co. (St. Louis, MO, USA); VCR were purchased from Shenzhen Wan Le Pharmaceutical Co., Ltd. (Shenzhen, Guangdong, China); cell culture media, fetal bovine serum (FBS) and other supplementary materials were purchased from Gibco Co. (Grand Island, NY, USA).

Techniques: Western Blot, BrdU Incorporation Assay, Flow Cytometry, Expressing, DNA Synthesis, Gene Expression, Transfection, Comparison, Control

Journal: The Journal of Biological Chemistry

Article Title: Bromodomain and Extraterminal Protein Inhibition Blocks Growth of Triple-negative Breast Cancers through the Suppression of Aurora Kinases *

doi: 10.1074/jbc.M116.738666

Figure Lengend Snippet: Aurora kinases are downstream targets of BETi. A, left panel, representative images (20×) of the indicated cell lines treated with vehicle or 500 nm JQ1 for 96 h and stained with DAPI (blue, nuclei) and Texas Red-X phalloidin (red, actin cytoskeleton). Insets show examples of multinucleated cells. Arrows indicate multinucleated cells. Bars, 50 μm. Right panel, quantitation of multinucleated cells. The data are means ± S.D. B, MDA-MB-231 cells were treated for 8 days, stained with propidium iodide, and analyzed by flow cytometry. The data are means ± S.E. (p < 0.05 compared with vehicle for both 2N and 4N populations). C, RT-qPCR analysis of AURKA and AURKB expression in the indicated TNBC cell lines treated with vehicle or 500 nm JQ1 for 24 h. The data are means ± S.D. D, Western blotting analysis of AURKA and AURKB expression in four TNBC cell lines treated with vehicle or 500 nm JQ1 for 24 h. The values on the Western blot are relative to untreated samples per cell line following normalization to β-actin. E, representative gene-specific ChIP-PCR analysis of MDA-MB-231 cells assessing binding of BRD4 to AURKA and AURKB. CDKN1A was used as a control that does not lose BRD4 binding with JQ1 treatment. Negative primers were designed to areas outside the promoter region for each gene. The data are means ± S.D. F, RT-qPCR analysis of AURKA and AURKB expression in MDA-MB-231 and MDA-MB-468 tumors from mice treated for 28 (MDA-MB-231 cells) or 35 (MDA-MB-468 cells) days with vehicle or JQ1. The data are means ± S.D. *, p < 0.05 compared with vehicle, for all graphs. (B), basal; (C), claudin low.

Article Snippet: Quantitative real time PCR was performed on an Applied Biosystems Step One Plus real time PCR system using the following TaqMan Gene Expression Assays (Thermo Fisher): 1) AURKA (Hs01582072_m1), 2) AURKB (Hs00945858_g1), 3) MYC (Hs00153408_m1), and 4) GAPDH (Hs02758991_g1).

Techniques: Staining, Quantitation Assay, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Binding Assay

Journal: iScience

Article Title: AURKB and PI3K/AKT/mTOR pathways converge to regulate TERT expression

doi: 10.1016/j.isci.2025.113194

Figure Lengend Snippet: Correlation and impact of AURKB expression on TERT levels in thyroid cancer subtypes (A) Thyroid cancer cells transfected with AURKB-specific shRNA plasmids were harvested following 4-day puromycin selection for assessment of AURKB levels by western blotting and TERT by RT-qPCR. Plotted values are the mean ± SD of three independent experiments, each performed in triplicate, expressed as fold-change in normalized expression relative to untransfected control. (B–F) Correlation between TERT and AURKB expression determined by RT-qPCR in TERTp mutated thyroid tumor cell lines: TPC1 (PTC), SW1736, 8505C, and C643 (ATC), alongside the non-malignant NThy-ori-3.1 cell line. Correlation between TERT and AURKB expression in (C) 220 PTC samples from the TCGA study, and (D) 37 advanced thyroid cancers from the Landa et al. study, using z-scores calculated from available transcriptomic data. Quantitative ChIP of AURKB, EZH2 and H3S10ph, H3K4Me3 and H3K27Me3 epigenetic marks at specific regions spanning the TERT transcriptional start site. Chromatin was prepared from (E) mutant C643 or TPC1, and (F) wild-type CAL62 and MDAT-T41 cells treated for 24 h with either 180 nM Hesperadin or DMSO. Plotted values are the mean average ± SD of three independent experiments. For all experiments, asterisks (∗) denote significant changes, determined at p < 0.05, using an unpaired two-tailed t -test with the assumption of equal variances.

Article Snippet: Human AURKB shRNA Plasmid Kit , OriGene , Cat#:TL320607.

Techniques: Expressing, Transfection, shRNA, Selection, Western Blot, Quantitative RT-PCR, Control, Mutagenesis, Two Tailed Test

Journal: iScience

Article Title: AURKB and PI3K/AKT/mTOR pathways converge to regulate TERT expression

doi: 10.1016/j.isci.2025.113194

Figure Lengend Snippet: Impact of AURKB inhibition on TERT expression and telomerase activity (A–C) Dose response of TERT expression measured by RT-qPCR across thyroid cancer cell-lines treated with 0.1–10 μM Hesperadin. Plotted values are the mean ± SD of three independent experiments, each performed in triplicate, expressed as fold-change in normalized expression relative to DMSO control. Time-course analysis showing modulation of (B) TERT expression 24–72 h measured by RT-qPCR and (C) telomerase activity relative to MCF7 control 72–120 h measured by qTRAP, following treatment with IC50 dose of Hesperadin across various cancer cell lines. (D–F), Cell-cycle independent effect of Hesperadin on TERT expression was assessed by RT-qPCR at 0, 3, and 6 h post IC50 Hesperadin dose in SW1736 and CAL62 cells, which were arrested at the G2/M phase by Nocodazole. Flow cytometry was utilized to evaluate the cell cycle distribution of cells exposed to a 24 h Nocodazole vehicle, 24 h Nocodazole with a 6 h Hesperadin vehicle, or Nocodazole plus 6 h Hesperadin treatment. Data illustrates average cell percentages in G1, S, and G2/M phases (±SD). Values presented are mean ± SD from three independent experiments, each conducted in triplicate, expressed as fold changes in normalized expression/activity relative to 0 h baseline levels. For all experiments, asterisks (∗) denote significant changes, determined at p < 0.05, using an unpaired two-tailed t -test with the assumption of equal variances.

Article Snippet: Human AURKB shRNA Plasmid Kit , OriGene , Cat#:TL320607.

Techniques: Inhibition, Expressing, Activity Assay, Quantitative RT-PCR, Control, Flow Cytometry, Two Tailed Test

Journal: iScience

Article Title: AURKB and PI3K/AKT/mTOR pathways converge to regulate TERT expression

doi: 10.1016/j.isci.2025.113194

Figure Lengend Snippet: mTORC1 inhibition leads to the dissociation of TRIM28-TRIM24 and AURKB from the mutant TERT promoter (A–F) Representative western blot images showing TRIM28 and TRIM24 expression across thyroid cell-lines. Co-immunoprecipitation of (B) endogenous TRIM24 in SW1736 and C643 cells; or (C) exogenous flag-tagged proteins overexpressed in SW1736 cells. Whole-cell lysates were incubated with anti-TRIM28 or IgG antibody for IP. (D, Left hand panel) Representative western blot images showing REST and expression across thyroid cell-lines. (D, Right hand panel) Co-immunoprecipitation of endogenous TRIM28 and AURKB by REST proteins in SW1736. Quantitative ChIP analysis was performed to measure TRIM28, TRIM24, AURKB, REST, EZH2 and H3S10ph levels at specific regions around the TERT transcriptional start site in (E) TPC1 and C643, and (F) CAL62 and MDAT-T41 cell following treatment with 2 nM Everolimus or DMSO for 6 h. The plotted values represent the mean ± SD derived from three independent experiments. In all experiments, asterisks (∗) indicate significant changes, determined at p < 0.05 using an unpaired two-tailed t -test with the assumption of equal variances.

Article Snippet: Human AURKB shRNA Plasmid Kit , OriGene , Cat#:TL320607.

Techniques: Inhibition, Mutagenesis, Western Blot, Expressing, Immunoprecipitation, Incubation, Derivative Assay, Two Tailed Test