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. 2010 Mar;17(1):73–85. doi: 10.1677/ERC-09-0100

Figure 4.

Figure 4

Pit-1 leads to a PRL-independent increase in cyclin D1. (A and C) Western blots of Pit-1, PRL, cyclin D1, and β-actin (used as loading control) were performed on control, pRc/RSV, and pRSV-hPit-1-transfected MCF-7 cells at 12 and 48 h respectively. (B and D) Relative protein levels in control MCF-7 cells and cells transfected with the pRc/RSV or pRSV-hPit-1 vector after 12 and 48 h respectively were calculated as the ratio of Pit-1, cyclin D1, and PRL with respect to the β-actin protein levels. Values represent mean±s.d. from densitometric evaluation of three western blots from independent experiments. (E) MCF-7 cells were transfected with 20 nM missense siRNA or 20 nM PRL siRNA. Forty-eight hours later, PRL protein immunoreactivity was evaluated by western blot. (F) MCF-7 cells were cotransfected with the pRSV-hPit-1 overexpression vector (2.5 μg) together with PRL siRNA (20 nM), and 48 h later, Pit-1, PRL, cyclin D1, and b-actin (as loading control) were evaluated by western blot.