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. 2006 Dec 18;175(6):937–946. doi: 10.1083/jcb.200604073

Figure 4.

Figure 4.

Rac1 and MgcRacGAP were required for IL-3–induced nuclear accumulation and transcriptional activation of p-STAT5A. (A) IL-3–induced transcriptional activation of STAT5A was suppressed by knock down of Rac1 or MgcRacGAP. Expression of bcl-xL or GAPDH mRNA was examined in Ba/F3 cells treated with the control siRNA (lane 1), Rac1 siRNA (lane 2), or MgcRacGAP siRNA (lane 3). 24 h after the siRNA treatment, the live cells were collected using Ficoll and subjected to semiquantitative RT-PCR. (B) IL-3–induced nuclear accumulation of p-STAT5A was impaired by knock down of Rac1 or MgcRacGAP. The intracellular distribution of p-STAT5A or total STAT5A in the IL-3–stimulated or unstimulated Ba/F3 cells pretreated with the control, Rac1, or MgcRacGAP siRNA (a and b). Note that Rac1 or MgcRacGAP expression was specifically suppressed by siRNA (c and d). Cytosol and nuclear extracts were prepared as described previously (Nakamura et al., 2002) and validated by Western blot using an anti-HDAC antibody or anti-RhoA antibody (e and f).