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. 2010 Jun 14;30(16):4006–4021. doi: 10.1128/MCB.01600-09

FIG. 7.

FIG. 7.

TBLR1 is required for the polyubiquitination of BCL-3. (A) Extended BCL-3 half-life in TBLR1-depleted cells. (Top) TBLR1-depleted Karpas cells were either left untreated (lanes 1 and 6) or stimulated with cycloheximide (CHX) (50 μg/ml) (lanes 2 to 5 and 7 to 10), and cell extracts were subjected to Western blotting (WB) with an antibody against Hsp90, BCL-3, or TBLR1, as indicated. (Bottom) Quantification of BCL-3 levels under control conditions or upon TBLR1 depletion in Karpas cells. The signal intensity in GFP siRNA-treated Karpas cells was set to 100%. (B) Impaired polyubiquitination (poly-Ub) of BCL-3 upon TBLR1 depletion. Karpas cells transfected with a siRNA against GFP or TBLR1 were stimulated with MG132. (Top two panels) Cell extracts were subjected to immunoprecipitation (IP) with an anti-FLAG (negative control) or anti-BCL-3 antibody, followed by Western blotting with an antibody against ubiquitin (Ub) or BCL-3. (Bottom three panels) Cell extracts were subjected to Western blotting with an antibody against BCL-3, Ub, or TBLR1 as well (bottom panels).