FIG. 2.

Turnover and subcellular localization of TRIM5α_rh B-box 2 variants. (A) HeLa cells expressing wild-type TRIM5α_rh and TRIM5α_rh ER/RE were treated with cycloheximide to block protein synthesis for a 5-h period in the absence or presence of the proteasome inhibitor MG115. Cells were harvested and lysed at 1-h intervals. Cell lysates containing equal amounts of total protein were analyzed by Western blotting with anti-HA and anti-actin antibodies. (B) HeLa or Cf2Th cells stably expressing the HA-tagged TRIM5α_rh variants were stained with an anti-HA antibody, followed by an anti-rat secondary antibody conjugated to Alexa 488. The TRIM5α_rh proteins are shown in green, and the nuclei are stained blue with DAPI.