Journal:

Article Title: The Arf-inducing Transcription Factor Dmp1 Encodes a Transcriptional Activator of Amphiregulin, Thrombospondin-1, JunB and Egr1

doi: 10.1002/ijc.24938

Figure Lengend Snippet: Dmp1 regulates target gene expression in a non-transformed murine alveolar epithelial cell line, C10. C10 cells were transfected with pFLEX1-Dmp1 vector or control vector and mRNA levels were quantitated 48 hours after transfection by real-time RT-PCR. (a) Induction of Areg, Tsp-1, JunB, Egr1, Bcl-3, Mbd1 and Adm genes by Dmp1. These gene inductions were nullified by Actinomycin D treatment, suggesting that Dmp1 increased the mRNA levels by transcriptional activation. (b) Repression of the Gas1 and Ect2 genes by Dmp1. This gene repression was also neutralized by Actinomycin D treatment, suggesting that the repression occurred at the transcriptional level. The p27kip1 gene, which did not show any significant change in the GeneChip Microarray, was not affected by Dmp1 overexpression in C10 cells. (c) Primary quantitative primary transcript real-time PCR (qPT-PCR) analysis on Areg, Tsp1 and Egr1 genes. The relative abundance of immature (pre) and mature Areg, Tsp-1 and Egr1 transcripts were similarly elevated at 24 and 48 hours in C10 cells transfected with the Dmp1 expression vector.

Article Snippet: Once the probe arrays were hybridized and washed, they were scanned using the Affymetrix GeneChip 7G plus scanner (Hewlett-Packard Gene array scanner G2500A; Affymetrix) according to the manufacturer’s procedures.

Techniques: Expressing, Transformation Assay, Transfection, Plasmid Preparation, Quantitative RT-PCR, Activation Assay, Microarray, Over Expression, Real-time Polymerase Chain Reaction