Higashi, K., Inagaki, Y., Suzuki, N., Mitsui, S., Mauviel, A., Kaneko, H. and Nakatsuka, I.
Notes: The Y-box-binding protein YB-1 was examined through the use of a reporter plasmid with wildtype and mutant putative Y-box binding sites. The binding sites were constructed in the pGL3 Basic Vector and measured using the Luciferase Assay System. Studies were performed in normal human dermal fibroblasts. Expression of YB-1 was found to repress expression of the COL1A2 gene at the transcriptional level. Confirmation of this dose-dependent inhibition was through measurement of the steady-state mRNA levels by quantitative, real-time RT-PCR. The reverse transcription portion of the quantitative, real-time RT-PCR reactions were performed with ImProm-II™ Reverse Transcriptase followed by a TaqMan-type quantitative PCR step. (2623)