Journal: PLoS ONE

Article Title: Bortezomib Reduces the Tumorigenicity of Multiple Myeloma via Downregulation of Upregulated Targets in Clonogenic Side Population Cells

doi: 10.1371/journal.pone.0056954

Figure Lengend Snippet: (A). Real time quantitative PCR of cell cycle and mitosis related genes ( CCNB1,CDC2, CDC20, CDC25C, AURKB, BIRC5, TOP2A, ASPM), Polycomb related genes ( EPC1, EZH2), and ubiquitin-proteasome related gene ( UBE2D3 and PSMA5) against RPMI 8226, AMO1, KMS-12-BM, JJN3 and KMS-11 cells. Y-axis: gray and white bars depict 2 −ΔΔCt values for gene expression. Asterisks (*) indicate statistical significance: *0.01≤ P <0.05, **0.001≤ P <0.01, *** P <0.001. Bars are means ± SD of three independent experiments. (B). Western blot analysis of Cyclin B1, CDC2, p-WEE1, p-CDC2, Aurora B, p-Aurora B, p-Hist.H3, EZH2, PSMA5 and GAPDH in SP and MP against RPMI 8226 and AMO1 cell lines.

Article Snippet: TaqMan probes of CCNB1 (Hs01030097_m1), EZH2 (Hs01016789_m1), TOP2A (Hs00172214_m1), CDC2 (Hs00938777_m1), CDC20 (Hs00415851_g1), CDC25C (Hs00156411_m1), ASPM (Hs00411505_m1), AURKB (Hs00177782_m1), BIRC5 (Hs00220565_m1), UBE2D3 (Hs00704312_m1), PSMA5 (Hs00936004_m1), EPC1 (Hs00228677_m1) and GAPDH (Hs02758991_g1) were purchased from Applied Biosystems.

Techniques: Real-time Polymerase Chain Reaction, Ubiquitin Proteomics, Gene Expression, Western Blot

Journal: PLoS ONE

Article Title: Bortezomib Reduces the Tumorigenicity of Multiple Myeloma via Downregulation of Upregulated Targets in Clonogenic Side Population Cells

doi: 10.1371/journal.pone.0056954

Figure Lengend Snippet: (A). SP of primary samples (M4, M7 and M8). Left panel: cells obtained through bone marrow aspiration gated for CD138 + with and without 50 µM reserpine. SP fractions (%) are shown beside the SP gates surrounded by black lines. (B). Real time PCR analysis of eight samples of MM primary tumor cells. Shown bar graphs are CCNB1, EZH2, AURKB and PSMA5 in SP and MP cells of indicated five myeloma cell lines. Y-axis: gray and white bars depict 2 −ΔΔCt values for gene expression. Asterisks (*) indicate statistical significance: *0.01≤ P <0.05, **0.001≤ P <0.01, *** P <0.001. Bars are means ± SD of triplicate samples.

Article Snippet: TaqMan probes of CCNB1 (Hs01030097_m1), EZH2 (Hs01016789_m1), TOP2A (Hs00172214_m1), CDC2 (Hs00938777_m1), CDC20 (Hs00415851_g1), CDC25C (Hs00156411_m1), ASPM (Hs00411505_m1), AURKB (Hs00177782_m1), BIRC5 (Hs00220565_m1), UBE2D3 (Hs00704312_m1), PSMA5 (Hs00936004_m1), EPC1 (Hs00228677_m1) and GAPDH (Hs02758991_g1) were purchased from Applied Biosystems.

Techniques: Real-time Polymerase Chain Reaction, Gene Expression

Journal: Frontiers in genetics

Article Title: Assessing the Potential Prognostic and Immunological Role of TK1 in Prostate Cancer.

doi: 10.3389/fgene.2022.778850

Figure Lengend Snippet: FIGURE 4 | Protein networks and the correlation between TK1 and the hub genes. (A,B) Molecular Complex Detection (MCODE) components of the hub genes. (C) The expression of serval hub genes was down-regulated in the TK1-silencing cells verified by RT-PCR. (D) TK1 and AURKB protein expression showed by immunohistochemical staining in the same high-grade and low-grade patient. The pictures were taken from the Human Protein Atlas dataset.

Article Snippet: For the PCa specimen shown in the figures, TK1 antibody (Atlas Antibodies, Cat# CAB004683) and AURKB antibody (Atlas Antibodies, Cat#CAB005862) were applied.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Immunohistochemical staining, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Maternally recruited Aurora C kinase is more stable than Aurora B to support mouse oocyte maturation and early development

doi: 10.1073/pnas.1120517109

Figure Lengend Snippet: AURKC is more stable than AURKB during meiotic maturation. (A–C) GV-intact oocytes were coinjected with the indicated cRNAs and matured to the indicated stage. In each graph, the first time point is 1 h after cycloheximide addition, and fluorescent images were obtained at the indicated times. Below are representative images from each time course. Data represent mean (± SEM) from at least 30 oocytes from two independent experiments. (Scale bars, 5 μm.) (D) Merge of the data from A–C.

Article Snippet: Taqman probes specific for Aurkb (Mm01718146_g1) and Aurkc (Mm03039428_g1) (Applied Biosystems) were used for gene expression detection and the comparative C t method was used to determine the difference in expression levels between stages.

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Maternally recruited Aurora C kinase is more stable than Aurora B to support mouse oocyte maturation and early development

doi: 10.1073/pnas.1120517109

Figure Lengend Snippet: AURKB stability does not depend upon its N terminus. (A) Schematic representation of AURKB and AURKC. KEN, A-, and D-boxes are indicated, and the conserved regions are shaded in light gray. (B–D) GV-intact oocytes were coinjected with the indicated cRNAs and matured to the indicated stage. Cycloheximide was added 1 h before the first time point and fluorescent images were obtained at the indicated times. Data represent mean (± SEM) from at least 30 oocytes from two independent experiments. MG132 was added as indicated to inhibit the proteasome (B).

Article Snippet: Taqman probes specific for Aurkb (Mm01718146_g1) and Aurkc (Mm03039428_g1) (Applied Biosystems) were used for gene expression detection and the comparative C t method was used to determine the difference in expression levels between stages.

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Maternally recruited Aurora C kinase is more stable than Aurora B to support mouse oocyte maturation and early development

doi: 10.1073/pnas.1120517109

Figure Lengend Snippet: Aurkc is a maternally recruited message. (A) Schematic representation of the Firefly luciferase (Luc) fusions to the Aurkb and Aurkc 3′ UTRs. Putative CPE and HEX motifs are underlined and the number of nucleotides between the motifs are indicated in the gray boxes. In Aurkc, the two nucleotides in the CPE that were mutated are in bold. Note that the lengths are not to scale. (B) GV-intact oocytes were coinjected with Luc fused to the indicated 3′ UTR and Renilla luciferase as an injection volume control. Luminescence was measured and quantified as the fold-difference in MII compared with GV. Data represent mean (± SEM) from 10 oocytes from two independent experiments. mut, mutated.

Article Snippet: Taqman probes specific for Aurkb (Mm01718146_g1) and Aurkc (Mm03039428_g1) (Applied Biosystems) were used for gene expression detection and the comparative C t method was used to determine the difference in expression levels between stages.

Techniques: Luciferase, Injection, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Maternally recruited Aurora C kinase is more stable than Aurora B to support mouse oocyte maturation and early development

doi: 10.1073/pnas.1120517109

Figure Lengend Snippet: Ectopic expression of AURKB in oocytes and embryos from Aurkc KO mice rescues their defects. (A and C) GV-intact oocytes or one-cell embryos from a single mouse of the indicated genotype were subdivided and microinjected with the indicated cRNAs and matured to MII to determine incidence of PBE or developed to the two-cell stage to monitor cytokinesis. These experiments were repeated three times with two to four KO mice per experiment. (B) Wild-type or Aurkc KO oocytes were microinjected with Aurkb-Gfp cRNA, matured to MI and MII, and imaged live. Shown are representative images. (Scale bar, 5 μm.) One-way ANOVA was used to analyze the data. *P < 0.05; mCh, mCherry.

Article Snippet: Taqman probes specific for Aurkb (Mm01718146_g1) and Aurkc (Mm03039428_g1) (Applied Biosystems) were used for gene expression detection and the comparative C t method was used to determine the difference in expression levels between stages.

Techniques: Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Maternally recruited Aurora C kinase is more stable than Aurora B to support mouse oocyte maturation and early development

doi: 10.1073/pnas.1120517109

Figure Lengend Snippet: Oocytes contain a third AURK to support oocyte maturation and early development because AURKB is not stable during meiosis and it is not recruited for translation as efficiently. Somatic cells continually cycle from G1 phase through mitosis during their life span. AURKB (orange) localizes to centromeres during metaphase, is destroyed at the end of the cell cycle, and is resynthesized during the next cell cycle. Oocytes contain both AURKB and AURKC (blue) (green where they colocalize). Both proteins (colored circles) are gradually degraded during meiosis, but AURKB does so more rapidly than AURKC does. There is no intervening cell cycle between meiosis I (MI) and meiosis II (MII) and oocytes are transcriptionally quiescent, making this cell type dependent on mRNA recruitment to synthesize adequate amounts of AURK. Aurkc message (colored lines) is recruited for translation more efficiently than Aurkb message is. The boxes around regions of chromosomes refer to the zoomed in Insets that contain cartoon labels representing AURKB and AURKC. A, anaphase; M, mitosis; S, synthesis; T, telophase; *, polar body.

Article Snippet: Taqman probes specific for Aurkb (Mm01718146_g1) and Aurkc (Mm03039428_g1) (Applied Biosystems) were used for gene expression detection and the comparative C t method was used to determine the difference in expression levels between stages.

Techniques: