FIGURE 5.

Elevated transcription from the Tsp-1 promoter in the Nedd4-1^−/−,trap MEFs. A, luciferase activity reporting Tsp-1 promoter activity in WT (+/+) or Nedd4-1^−/−,trap (−/−) MEFs. The Tsp-1 promoter was expressed from the pGL3 vector in the presence/absence of the indicated amounts of c-Myc cDNA. Shown is a representative experiment, performed in duplicate. **, p ≤ 0.01, unpaired t test. In three separate experiments, each performed in duplicate, there was a 3.3-fold increase in Tsp-1 transcription in the Nedd4-1^−/−,trap relative to WT MEFs (p = 3 × 10⁻⁹, unpaired t test). B, semi-quantitative PCR (sqPCR) revealing increased mRNA levels in Nedd4-1^−/−,trap MEFs relative to WT. Quantification of mRNA levels, analyzed by quantitative PCR, revealed 3.98-fold increase in mRNA of Tsp-1 in the Nedd4-1^−/−,trap MEFs relative to WT MEFs.