Skip to main content
. 2005 Nov 28;33(20):6528–6539. doi: 10.1093/nar/gki956

Figure 2.

Figure 2

Interaction among Sp1, p621 and Rta in vitro. GST–p621 was added to the lysate prepared from P3HR1 cells treated with TPA and sodium butyrate. Proteins bound to GST–p621 were pulled down with glutathione–Sepharose beads (Beads) and analyzed by IB using anti-Rta (A) and anti-Sp1 antibody (B). Parallel experiments involving GST–Rta (lane 3) and GST–Sp1 (lane 4) were also conducted and IB was performed with anti-MCAF1 antibody (C). Meanwhile, anti-HA antibody (lanes 2 and 3) and anti-Sp1 antibody (lanes 4 and 5) were added to mixtures containing Sp1, GST–p621 and Rta to determine whether the interaction between Sp1 and Rta requires p621 (D). Various amounts of Rta were added to binding mixtures that contained Sp1 and GST–p621. Proteins were pulled down using GST–p621–glutathione–Sepharose beads and analyzed by IB with anti-Sp1 antibody (E). In each part, lane 1 was loaded with cell lysate; in lane 2, proteins were eluted from GST–glutathione–Sepharose beads.