Journal: Molecular Cancer

Article Title: The role of high cell density in the promotion of neuroendocrine transdifferentiation of prostate cancer cells

doi: 10.1186/1476-4598-13-113

Figure Lengend Snippet: Androgen depletion and high cell density both induce cell cycle arrest in LNCaP and LAPC-4 cells. A , Analysis of changes in cell cycle distribution in response to high cell density (FBS) and androgen depletion (CS). The data represent means ± SD of three independent experiments. “*”denotes statistical significance compared with control (2 days in FBS), “ # ” denotes statistical significance compared with 2 days in CS. B , Western blot analysis of the expression of selected cell cycle regulators in LNCaP and LAPC-4 cells. Graphs represent optical density (O.D.) of p-Cdk2 normalized to O.D. of total Cdk2.

Article Snippet: LNCaP and LAPC-4 cells were transfected with small inhibitory RNA (siRNA) duplexes (Santa Cruz Biotechnology) directed against non-targeting control (sc-37007), Cdk1 siRNA (sc-29252), and Cdk2 siRNA (sc-156139) using the Neon® Transfection System (Life Technologies).

Techniques: Western Blot, Expressing

Journal: Molecular Cancer

Article Title: The role of high cell density in the promotion of neuroendocrine transdifferentiation of prostate cancer cells

doi: 10.1186/1476-4598-13-113

Figure Lengend Snippet: Deregulation of the cell cycle by inhibition of Cdk1 and/or Cdk2 boosts expression of NED markers in AR-positive prostate cancer cell lines. A-D, LAPC-4 cells were treated with an inhibitor of Cdk2 activity, CVT-313 (10 μM, 48 h). A , Western blot analysis of Rb protein. B , Analysis of cell cycle distribution. The data represent means ± SD (n=4). C , Western blot analysis of expression of γ-enolase and tubulin β-III. D , qRT-PCR analysis of TUBB3, CHGA, and ENO2 mRNA level. The data represent means ± SD (n=4). “*” denotes statistical significance compared with control. E - G , LAPC-4 cells were transfected with control siRNA or with a combination of Cdk1- and Cdk2 siRNAs for 48 hours. E , Western blot analysis of Cdk1 and Cdk2 48 hours after transfection. F , Analysis of cell cycle distribution in response to Cdk1 and Cdk2 down-regulation. The data represent means ± SD (n=3). G , qRT-PCR analysis of changes in mRNA levels of TUBB3 and CHGA in response to Cdk1 and Cdk2 down-regulation. The data represent means ± SD (n=4). “*”denotes statistical significance compared with control siRNA. H - J , LNCaP and LAPC-4 cells were treated with DMSO or the indicated Cdk1 inhibitors for 48 hours (LNCaP: 2 μM CGP, 7.5 μM RO; LAPC-4: 3.5 μM CGP, 7.5 μM RO). H , Analysis of cell cycle distribution in LNCaP and LAPC-4 cells treated with the indicated Cdk1 inhibitors. The data are presented as means ± SD of one out of two independent experiments performed in duplicate. I , Western blot analysis of γ-enolase and tubulin β-III in response to Cdk1 inhibitors. J , qRT-PCR analysis of TUBB3 and ENO2 in response to Cdk1 inhibitors. The data are presented as means ± SD of one out of two experiments performed in duplicate. CGP, CGP 74514A; RO, RO-3306.

Article Snippet: LNCaP and LAPC-4 cells were transfected with small inhibitory RNA (siRNA) duplexes (Santa Cruz Biotechnology) directed against non-targeting control (sc-37007), Cdk1 siRNA (sc-29252), and Cdk2 siRNA (sc-156139) using the Neon® Transfection System (Life Technologies).

Techniques: Inhibition, Expressing, Activity Assay, Western Blot, Quantitative RT-PCR, Transfection