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. 2011 Jan 3;286(9):7609–7618. doi: 10.1074/jbc.M110.166603

FIGURE 5.

FIGURE 5.

Identification of DUPD1 as PRL-RS associated protein. PRL-RS-GST (RS-GST) fusion proteins were prepared as described under “Experimental Procedures.” RS-GST interacting proteins were pulled down using decidual cell extracts from PRLR−/−RS mice treated with PRL (60 μg/animal) or vehicle (A) or GG-CL cells transfected with PRL-RS and treated with PRL (1 μg/ml) for different time points (B). The bands were detected by silver staining. The molecular mass markers (lanes M) were as indicated to determine the size of the bands. As a control, a well of pre-pulldown PRL-RS-GST (RS) was loaded showing a band of 25 kDa. The band (27–30 kDa) pulldown from decidual and GG-CL cell lysates by RS-GST fusion protein is indicated by arrow. C, RS-GST pulldown samples of GG-CL cell lysates were analyzed by Western blot using a polyclonal antibody to DUPD1. To show the specific interaction of DUPD1 with RS-GST, GST alone (C-GST) was used to pull down GG-CL cell lysates as a control. GG-CL cells were transfected with PRL-RS and treated with PRL (1 μg/ml) for different time points. Expression of DUPD1 was measured by qPCR (D) or Western analysis (E). *, p < 0.05 versus 0 min.