Journal: Investigational New Drugs

Article Title: The kinesin Eg5 inhibitor K858 exerts antiproliferative and proapoptotic effects and attenuates the invasive potential of head and neck squamous carcinoma cells

doi: 10.1007/s10637-022-01238-2

Figure Lengend Snippet: A – Immunofluorescence of cyclin B1 in untreated SCC-15 cells ( a ) and SCC-15 cells treated with 1 μM K858 for 24 h shown with ( b ) and without ( c ) nuclear staining. Bar, 7 μm. B – Western blot of cyclin B1 and tubulin in untreated (CTR) HNSCC cells and HNSCC cells treated with 1 μM K858 for 24 h

Article Snippet: Alternatively, after fixation, the cells were permeabilized with 0.1% Triton for 10 min at RT, incubated with 3% bovine serum albumin (BSA) for 1 h at RT and then incubated at 4 °C overnight with a primary rabbit polyclonal antibody to cyclin B1 with 0.1% BSA (1:200; Elabscience, TX, USA).

Techniques: Immunofluorescence, Staining, Western Blot

Journal: Journal of Virology

Article Title: The ATM/ATR Signaling Effector Chk2 Is Targeted by Epstein-Barr Virus Nuclear Antigen 3C To Release the G 2 /M Cell Cycle Block

doi: 10.1128/jvi.00053-07

Figure Lengend Snippet: FIG. 3. Nocodazole suppresses the phosphorylation of Cdc2 at Tyr15. (A) Western blot analysis of G2/M cell cycle regulators cyclin B1 and Cdc2 in BJAB control cells and EBNA3C (E3C)-transfected BJAB cells following exposure to nocodazole. Stably transfected cells were treated with nocodazole (200 ng/ml) for 24 h and then harvested, and total cell lysates were prepared. Following electrophoresis and transfer onto membranes for Western blot analysis, the membranes were probed with antibodies () for cyclin B1, total Cdc2, Tyr15-phosphorylated Cdc2 [Cdc2(Tyr15)], and -actin as a protein control. , present; , absent. (B) Results of in-cell Western analysis showing the phosphorylation at Tyr15 of Cdc2 in the presence of EBNA3C. BJAB cells carrying the vector control or stably transfected with EBNA3C were grown in round-bottom 96-well plates. The cells were then treated with nocodazole. The results of in-cell Western analysis for Tyr15 phosphorylation are presented with respect to the basal level of Cdc2 and are expressed as relative intensities. Means and standard deviations were derived from three independent experiments.

Article Snippet: Polyclonal rabbit anti-phospho-Tyr15 Cdc2 (catalog no. 9111), rabbit polyclonal anti- -actin, mouse monoclonal Cdc2 (catalog no. 9116), mouse monoclonal cyclin B1, and rabbit polyclonal Cdc25c antibodies were all purchased from Cell Signaling Inc. (Danvers, MA).

Techniques: Phospho-proteomics, Western Blot, Control, Transfection, Stable Transfection, Electrophoresis, In-Cell ELISA, Plasmid Preparation, Derivative Assay

Journal: Journal of Virology

Article Title: The ATM/ATR Signaling Effector Chk2 Is Targeted by Epstein-Barr Virus Nuclear Antigen 3C To Release the G 2 /M Cell Cycle Block

doi: 10.1128/jvi.00053-07

Figure Lengend Snippet: FIG. 9. A hypothetical model shows the putative mechanisms for the bypassing of the nocodazole-induced G2 arrest by EBNA3C (E3C). Nocodazole treatment reduces the level of phosphorylated Cdc2. The viral nuclear antigen EBNA3C binds directly to Chk2, which results in the phosphorylation of Cdc25c at Ser216. Cdc25c, which is phosphorylated predominantly at Ser216, is sequestered in the cytoplasm and is now unable to regulate the phosphorylation of nuclear Cdc2. The resulting effect leads to the activation of cyclin B-Cdc2 and progression through the G2/M stage, releasing the block imposed by nocodazole. The overall effect is the release of the nocodazole-induced G2 arrest.

Article Snippet: Polyclonal rabbit anti-phospho-Tyr15 Cdc2 (catalog no. 9111), rabbit polyclonal anti- -actin, mouse monoclonal Cdc2 (catalog no. 9116), mouse monoclonal cyclin B1, and rabbit polyclonal Cdc25c antibodies were all purchased from Cell Signaling Inc. (Danvers, MA).

Techniques: Phospho-proteomics, Activation Assay, Blocking Assay

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