Skip to main content
Figure 7 | BMC Molecular Biology

Figure 7

From: Thrombin induces Egr-1 expression in fibroblasts involving elevation of the intracellular Ca2+ concentration, phosphorylation of ERK and activation of ternary complex factor

Figure 7

Transcriptional upregulation of Egr-1 promoter/luciferase reporter gene transcription in thrombin or carbachol stimulated 39M1-81 cells. (A) Schematic representation of integrated proviruses encoding Egr-1 promoter/luciferase reporter genes. The transfer vectors pFWEgr1.2luc (# 1) and pFWEgr-1.1luc (# 2) contain the sequences from -490 to + 235 or from -239 to +235 derived from the human Egr-1 gene. The transfer vector pFWEgr-1SREluc (# 3) contains the two proximal SREs # 1 and 2 of the Egr-1 promoter upstream of a minimal promoter. The important genetic elements within the Egr-1 regulatory region are depicted, including five serum response elements (SRE), and a cAMP response element (CRE). The U3 region of the 5' LTR of the transfer vector is deleted. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) and the HIV flap element are indicated. (B, C) 39M1-81 cells (B) or 39M1-81-ΔRaf-1:ER cells (C) were infected with recombinant lentiviruses generated with the transfer vectors pFWEgr-1.1luc, pFWEgr-1.2luc, or pFWEgr-1SREluc. The infected cells were treated with thrombin, carbachol, or 4OHT for sixteen hours. Cell extracts were prepared and analyzed for luciferase activities. Luciferase activity was normalized to the protein concentration. Data are means ± SD for n = 4 experiments.

Back to article page