Figure 2
From: Enterovirus 71 induces degradation of TRIM38, a potential E3 ubiquitin ligase
TRIM38 functions as an E3 enzyme and ubiquitinates itself ex vivo. (A) TRIM38 catalyzes the ubiquitination of cellular proteins. 293T cells were transfected with plasmids expressing Flag-TRIM38 (lanes 1 and 2), pRK5-HA-Ubiquitin (lanes 2 and 3). Whole cell lysates were subjected to Western blot analysis with antibodies against HA, Flag and β-actin. (B) TRIM38 is self-ubiquitinated. Similar to (A), except that cell lysates were immunoprecipitated with anti-Flag. Immunoprecipitates and aliquots of cell lysates were subjected to Western blot analysis with antibodies against HA, Flag, and β-actin. (C) TRIM38 mediates itself K48-ubiquitination. 293T cells were transfected with plasmids expressing Flag-TRIM38 (lanes 1 and 3), pRK5-HA-ubiquitin-K48 (lanes 2 and 3). Cell lysates were immunoprecipitated with anti-Flag or anti-HA. Immunoprecipitates and aliquots of cell lysates were subjected to Western blot analysis with antibodies against HA, Flag and β-actin.