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. 2008 Dec 5;283(49):34273–34282. doi: 10.1074/jbc.M802527200

FIGURE 1.

FIGURE 1.

HBZ promotes proteasomal degradation of c-Jun. A, HEK-293T cells were transfected with 0.5 μg of pcDNA3-HA-c-Jun and 3 μg of either pcDNA3 or pcDNA3-FLAG-HBZ. After 12 h, the cells were treated with or without MG132 for 12 h. Cell lysates were immunoblotted with the antibodies indicated. B, HEK-293T cells were transfected with 5 μg of pCAG-FLAG-HBZ or control vector. After 36 h, the cells were treated with cycloheximide (50 μg/ml) and collected at the indicated times. Cell lysates were analyzed by immunoblot analysis. C, upper panel: C8166 cells were transfected using lentiviruses transcribing short hairpin RNAs against HBZ or a nonspecific (n/s) sequence. The relative mRNA expression of HBZ, c-Jun or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was evaluated by semiquantitative reverse transcription-PCR 7 days after transfection. Efficiencies of lentivirus vector transfection, which were determined by green fluorescent protein expression, were >95%. Lower panel: C8166 cells were transfected as indicated in the upper panel. After 7 days, the cells were treated with cycloheximide (50 μg/ml) and collected at the indicated times. Cell lysates were analyzed by immunoblot analysis.