Journal: International Journal of Oncology
Article Title: Association and regulation of protein factors of field effect in prostate tissues
Figure Lengend Snippet: (A) Computational analysis of the EGR-1 recognition sequence [GCG(G/T)GGCG] in the genomic sequence 1,500 bp upstream and 500 bp downstream of the transcription initiation site of PDGF-A, MIC-1, and FASN. Black vertical lines and black rectangular boxes denote genomic sequences and exons, respectively; vertical arrow heads indicate EGR-1 recognition sequences. (B) EGR-1, PDGF-A, MIC-1, and FASN protein expression in RWPE-1 cells transiently transfected with pcDNA3.1/EGR-1 (EGR-1 overexpression) or pLKO.1/EGR-1 shRNA (EGR-1 suppression), and their empty plasmid controls. Double bands in EGR-1 represent post-translational modifications ( 44 ). The fold change difference compared to empty plasmid control and determined by densitometry as a ratio with β-actin signal is indicated in the small bar graphs (left bar, EGR-1 overexpression; right bar, EGR-1 suppression). (C) Relative mRNA expression of PDGF-A, MIC-1, and FASN in RWPE-1 cells transiently transfected with pcDNA3.1/EGR-1 (EGR-1 overexpression) or pLKO.1/EGR-1 shRNA (EGR-1 suppression), and their empty plasmid controls. Bars represent averages of triplicates ± standard deviation; * Statistical significance (p≤0.05) from pcDNA3.1 and pLKO.1 plasmid vector control, respectively. EGR-1, early growth response-1; PDGF-A, platelet-derived growth factor-A; MIC-1, macrophage inhibitory cytokine-1; FASN, fatty acid synthase.
Article Snippet: Trypsin-EDTA at 0.25% was used to detach the cells for splitting and reculturing. pcDNA3.1 control and pcDNA3.1/EGR-1 plasmids were a kind gift of Dr W. Xiao (University of Science and Technology of China, Hefei, China). pLKO.1 control and pLKO.1/EGR-1 shRNA plasmids were from Sigma (St. Louis, MO, USA).
Techniques: Sequencing, Genomic Sequencing, Expressing, Transfection, Over Expression, shRNA, Plasmid Preparation, Standard Deviation, Derivative Assay