Journal: International journal of oncology

Article Title: Atrazine promotes RM1 prostate cancer cell proliferation by activating STAT3 signaling.

doi: 10.3892/ijo.2016.3433

Figure Lengend Snippet: Figure 3. Atrazine accelerates the cell cycle in vitro. (A) PI staining and flow cytometry analysis. (B) Flow cytometry analysis comparing cell numbers between control and atrazine treated cells over a period of 48 h. Mean ± SEM of three independent experiments are presented. (C) qRT‑PCR analysis of p53, p21, cyclin D1 and cyclin B1 mRNA levels, **P<0.01. (D) Western blot analysis of P53, Cyclin D1 and Cyclin B1 protein amounts. (E) The P53, Cyclin D1 and Cyclin B1 protein levels from three independent experiments. Columns, mean; bars, SE (*P<0.05, **P<0.01 versus control).

Article Snippet: Anti-Grim-19, Stat3, MMP9, MMP2, VEGF, PCNA, P53, c-myc, Cyclin B1, Cyclin D1, Bcl-2, Bax, and Caspase-3 primary antibodies, as well as secondary antibodies were acquired from Proteintech Group (USA).

Techniques: In Vitro, Staining, Flow Cytometry, Control, Western Blot

Journal: International journal of oncology

Article Title: Atrazine promotes RM1 prostate cancer cell proliferation by activating STAT3 signaling.

doi: 10.3892/ijo.2016.3433

Figure Lengend Snippet: Figure 4. Atrazine accelerates the cell cycle in vivo. (A) qRT‑PCR analysis of p53, p21, cyclin D1 and cyclin B1 mRNA expression. (B) Western blot analysis of P53, P21, Cyclin D1 and Cyclin B1 protein amounts. (C) Quantitation of P53, P21, Cyclin D1 and Cyclin B1 protein levels from three independent experiments. Columns, mean; bars, SE (*P<0.05, **P<0.01 versus control).

Article Snippet: Anti-Grim-19, Stat3, MMP9, MMP2, VEGF, PCNA, P53, c-myc, Cyclin B1, Cyclin D1, Bcl-2, Bax, and Caspase-3 primary antibodies, as well as secondary antibodies were acquired from Proteintech Group (USA).

Techniques: In Vivo, Expressing, Western Blot, Quantitation Assay, Control

Journal: bioRxiv

Article Title: A truncated form of the p27 CDK inhibitor translated from pre-mRNA causes cell cycle arrest at G2 phase

doi: 10.1101/2022.01.12.476115

Figure Lengend Snippet: (A-D) Two hours after release from a double thymidine block (DTB), synchronized HeLa S3 cells were treated with MeOH or the indicated concentrations of Pla-B. Black triangles indicate the time points of cell harvest and sample preparation (A). Cell cycle was analyzed at the indicated time points by cytometry (B). Morphology of the cells was observed under a microscope and round cells were counted at the indicated time points (C). Protein samples were prepared at the indicated time points. The protein levels of indicated proteins and phosphorylation status of Cdk1 were analysed by immunoblotting. Protein levels of α-tubulin were analysed as an internal control (D). Error bars indicate s.d. (n = 3).

Article Snippet: The in vitro kinase assay was performed using the Cyclin A2/Cdk1 Kinase Enzyme System (Promega, Madison, WI, USA), recombinant Cyclin B1/Cdk1 (SignalChem Biotech Inc., Richmond, BC, Canada), and ADP-Glo kinase assay (Promega), according to the manufacturers’ instructions.

Techniques: Blocking Assay, Sample Prep, Cytometry, Microscopy, Western Blot

Journal: bioRxiv

Article Title: A truncated form of the p27 CDK inhibitor translated from pre-mRNA causes cell cycle arrest at G2 phase

doi: 10.1101/2022.01.12.476115

Figure Lengend Snippet: (A) Cells expressing Flag-p27* under the control of tetracycline were synchronized by a double thymidine block. The cells were treated with 1 μg/ml DOX at the same time as release from the double thymidine block. The cells were harvested at 8 h after release from the double thymidine block (G2/M phase) and then immunoprecipitation was performed using anti-DDDDK (Flag) antibodies. Flag-tagged and coimmunoprecipitated proteins were analysed by immunoblotting. (B, C) Purified Flag-tagged proteins were applied to an in vitro kinase assay reaction and kinase activities of Cyclin A2/Cdk1 (B) and Cyclin B1/Cdk1 (C) were measured. Statistical significance was assessed by the one-way ANOVA and Dunnett’s test (*: P < 0.05; **: P < 0.01; ***: P < 0.01). Error bars indicate s.d. (n = 3).

Article Snippet: The in vitro kinase assay was performed using the Cyclin A2/Cdk1 Kinase Enzyme System (Promega, Madison, WI, USA), recombinant Cyclin B1/Cdk1 (SignalChem Biotech Inc., Richmond, BC, Canada), and ADP-Glo kinase assay (Promega), according to the manufacturers’ instructions.

Techniques: Expressing, Blocking Assay, Immunoprecipitation, Western Blot, Purification, In Vitro, Kinase Assay