Deubiquitination of type 2 iodothyronine deiodinase by von Hippel–Lindau protein–interacting deubiquitinating enzymes regulates thyroid hormone activation
J. Clin. Invest. Cyntia Curcio-Morelli, et al. 112:189 doi:10.1172/JCI18348 [
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Figure 3Stabilization 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 Na
2[
75Se]O
3 for 16 hours and pulse-chased with 100 nM Na
2SeO
3; 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.
