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. 2001 Dec 1;15(23):3088–3103. doi: 10.1101/gad.944801

Figure 2.

Figure 2

LEF1 is SUMO-modified in vivo. (A) PIASy associates with SUMO-modified LEF1. LEF1 and T7-PIASy were transiently expressed in 293T cells. Equivalent amounts of total cellular protein were immunoprecipitated with anti-T7 mAb. Coimmunoprecipitated endogenous, SUMO-conjugated proteins were detected by an anti-SUMO2/3 immunoblot (upper left). The anti-SUMO2/3 immunoblot was stripped and reprobed with an anti-LEF1 antibody (upper right). Numbers to the right indicate the molecular mass of protein markers in kilodaltons. The expression of LEF1 or T7-PIASy proteins in total cell lysates was determined by anti-LEF1 (middle) or anti-T7 (bottom) immunoblots, respectively. (B) LEF1 is SUMO-modified in vivo. LEF1, T7-PIASy, and Flag epitope-tagged SUMO1 or SUMO2 were transiently expressed in 293T cells. Equivalent amounts of total cellular protein were immunoprecipitated with anti-T7 mAb. Coimmunoprecipitated LEF1 proteins were detected by an anti-LEF1 immunoblot (top panel). The expression of LEF1 or T7-PIASy proteins in total cell lysates was determined by anti-LEF1 (middle) or anti-T7 (bottom) immunoblots, respectively. (C) LEF1, Flag epitope-tagged SUMO1 or SUMO2 and T7-PIASy were transiently expressed in 293T cells. LEF1 proteins in total RIPA buffer cell lysates were detected by an anti-LEF1 immunoblot (top). The expression of T7-PIASy proteins in total cell lysates was determined by an anti-T7 immunoblot (bottom).