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. 2003 Jul 15;112(2):189–196. doi: 10.1172/JCI18348

Figure 3.

Figure 3

Stabilization of 75Se-D2 is due to decreased Ub-D2. (a) HEK-293 cells transiently coexpressing FLAG-wtD2 and/or VDU1 or VDU2 were labeled with Na2[75Se]O3 for 16 hours and pulse-chased with 100 nM Na2SeO3; cells were harvested at 0 or 2 hours. (b) Same as in a except that wtD1 was coexpressed with VDU1 or VDU2, with a chase time of 6 hours. Cell lysates in a and b were processed for immunoprecipitation with anti-FLAG antibody, and pellets were resolved by SDS-PAGE. The densitometric analysis is presented for each gel. (c) HEK-293 cells transiently expressing FLAG-CysD2, HA-Ub, and VDU1 or VDU2 were lysed and processed for immunoprecipitation with anti-FLAG antibody. The pellets were resolved by SDS-PAGE and probed with anti-HA antibody by Western analysis. HEK-293 cells transiently coexpressing FLAG-wtD2 and/or VDU1 or VDU2 were labeled with 75Se for 16 hours, and the cell lysates were processed as described in a. The corresponding immunoprecipitated nonubiquitinated 75Se-D2 is shown below each lane of 75Se-Ub-D2. Each experiment in a–c was performed twice.