Skip to main content
. 2010 Aug 30;107(37):16119–16124. doi: 10.1073/pnas.1004653107

Fig. 4.

Fig. 4.

PIAS1 activity is regulated by SF2/ASF. (A) SF2/ASF enhances PIAS1 activity on SUMO conjugation. HEK 293T cells were transfected with HA-SUMO3 (500 ng) and FLAG-PIAS1, T7-SF2/ASF or T7-SF2/ASF ΔRRM2 (500 ng each) as indicated, together with 50 ng of pECFP for transfection efficiency and loading control (total amount of DNA, 2 μg). Cells were lysed in Laemmli sample buffer after 48 h. Western blotting was performed with the antibodies indicated at the bottom of each panel. (B) HEK 293T cells were transfected with HA-SUMO1 (500 ng), FLAG-PIAS1 (100 ng), T7-SF2/ASF, or its deletion mutants (500 ng each) as indicated (total amount of DNA, 2 μg). Cells were lysed in Laemmli sample buffer after 48 h. Western blotting was performed with the antibodies indicated at the bottom of each panel. (C) SF2/ASF is critical for PIAS1-mediated SUMO conjugation. HEK 293T cells were transfected either with a control siRNA (siRNA Ctl) or an SF2/ASF specific siRNA. After 24 h, cells were retransfected with the plasmids expressing HA-SUMO3 and FLAG-PIAS1 (500 ng). Forty-eight hours later, cells were lysed in Laemmli sample buffer and subject to Western blot, as indicated at the bottom of each panel.