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. 2002 Feb;70(2):998–1001. doi: 10.1128/iai.70.2.998-1001.2002

FIG. 1.

FIG. 1.

In vivo deamidation (A) and polyamination (B) of Rac and Cdc42 by DNT. (A) C3H10T1/2 cells expressing FLAG-tagged Rac1 or Cdc42 were treated with DNT at 5 ng/ml for 20 h and subjected to immunoprecipitation with anti-FLAG antibody. The precipitates were subjected to SDS-PAGE followed by immunoblot analysis with anti-63E antibody. The arrows and asterisk indicate the deamidated GTPases and nonspecifically reactive bands, respectively. (B) C3H10T1/2 cells expressing the FLAG-GTPases were preloaded with [14C]putrescine and treated with DNT at 10 ng/ml for 24 h. The cell lysates or the immunoprecipitated (IP) FLAG-GTPases were subjected to SDS-PAGE followed by autoradiography. Lane 2 shows that several kinds of endogenous proteins were 14C-polyaminated in the DNT-treated cells. The arrow indicates 14C-polyaminated FLAG-Rac1 and FLAG-Cdc42.