Journal: PLoS ONE

Article Title: Genistein Affects Histone Modifications on Dickkopf-Related Protein 1 (DKK1) Gene in SW480 Human Colon Cancer Cell Line

doi: 10.1371/journal.pone.0040955

Figure Lengend Snippet: pCMV vector containing human DKK1 gene was used to transfect cells before sample collection for analysis. Empty pCMV vector was used as the control for the overexpression experiments. Three independent cell samples were analyzed and presented as the mean ± SEM. A ) mRNA expression of DKK1 and Cyclin D1 in regular and DKK1 -transfected SW480 cells. The mRNA expression was analyzed by RT-PCR. Data were normalized to internal control L7a. B ) DKK1 protein expression in regular cells and transfected SW480 cells. Whole cell protein extracts were collected from transfected SW480 cells and western blot analysis of DKK1 was performed as described in materials and methods. A representative blot is shown and the quantification represents the mean± SEM from 3 independent dishes. Actin was used as the loading control. C ) Cell cycle analysis of regular and DKK1 -transfected SW480 cells. Result was obtained by flow cytometry. Y-axis represents % of gated cells and x-axis shows different cell cycle stages, including G1, S and G2/M. D ) WST-1 proliferation assay in vector- and DKK1 -transfected SW480 cells. WST-1 signals were converted to actual cell numbers using a standard generated by serial dilutions of a known number of cells. Data were normalized to control for genistein treatment and vector for DKK1 transfection, respectively. Asterisks (*) indicate statistical significance compared with the respective control group (genistein to control; DKK1 to vector; p <0.05). For the knockdown of DKK1, siRNA duplexes against human DKK1 gene were transfected into SW480 cells. Scrambled sequences were used as the control siRNA. E ) mRNA expression of DKK1 and Cyclin D1 in control siRNA (N/S si) and DKK1 siRNA (DKK1 si)-treated SW480 cells by control and genistein treatments. The mRNA expression was analyzed by RT-PCR. Data were normalized to internal control L7a. F ) WST-1 proliferation assay in control siRNA and DKK1 siRNA SW480 cells. WST-1 signals were converted to actual cell numbers using a standard generated by serial dilutions of a known number of cells. Asterisks (*) indicate statistical significance compared to Control 0 µmol/L genistein ( p <0.05). The bracket indicates statistical difference between the N/S si and DKK1 si groups treated with genistein (p = 0.002).

Article Snippet: At the time of transfection, 300 nmol/L of siRNA against human scramble sequence (N/S si, QIAGEN, Valencia, CA) or 300 nmol/L of siRNA against human DKK1 ( DKK1 si, Integrated DNA Technologies) was prepared in 200 μL serum-free media (SFM).

Techniques: Plasmid Preparation, Control, Over Expression, Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Cell Cycle Assay, Flow Cytometry, Proliferation Assay, Generated, Knockdown

Journal: Oncotarget

Article Title: The role of transcriptional coactivator TAZ in gliomas

doi: 10.18632/oncotarget.12625

Figure Lengend Snippet: A. Expression of TAZ in LN229 cell was knocked down after Luc-TAZ si-Lentivirus transfection detected by Western blotting and RT-PCR. B. TAZ expression was suppressed in tumor samples of Lenti-TAZ si group detected by IHC. C. TAZ si induced apoptosis in xenograft specimen detected by TUNEL method. D. BLI signals in TAZ si xenografts were weaker than those in Lenti-NC xenografts. E. The survival of six mice in TAZ si group (39, 47, 54, 55, 59, 60 days, respectively) was longer than that in Lenti-NC group (27, 31, 39, 45, 46, 48 days, respectively), the survival of mice in lenti-TAZ si group was significantly prolonged ( p <0.05).

Article Snippet: The 6-week-old male balb/c nude mice were randomly divided into Lentivirus with TAZ siRNA (Lenti-TAZ si) and Lentivirus with scramble sequence (Lenti-NC) (Genepharma, China) groups with each group being consisted of six mice.

Techniques: Expressing, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, TUNEL Assay