Skip to main content
. 2000 Jul 18;97(15):8507–8512. doi: 10.1073/pnas.97.15.8507

Figure 3.

Figure 3

Elongin binding stabilizes VHL products. VHL expression vectors were transiently transfected into 293T cells, either with no cotransfection (left panels, − elongins BC) or with cotransfection of both elongin B and elongin C vectors (right panels, + elongins BC). Transfected cells were treated with cycloheximide (CHX) for various time points (indicated above each blot) or with lactacystin (L). Whole cell lysates were normalized for equal protein loading (10 μg) in each lane. VHL Western blotting was performed by using mAbs to either Flag (A and B) or VHL (11E12, C and D). Positions of Flag-pVHL (A and B, Upper panels) are indicated by arrowheads to the left of each blot. Positions of untagged VHLp24(MPR) proteins (C and D, Upper panels) are indicated by a bracket to the left of each blot. Position of a faster-migrating VHLp24(MPR) polypeptide in C is indicated by an arrowhead to the right of the blot. Elongin B-HA and elongin C-Flag (A–D, Lower panels), detected with antibodies to the appropriate epitope-tag, also are indicated by arrowheads. VHL constructs used: Flag-VHLp24(MPR) (A); Flag-VHLp24(RC161/2QW) (B); VHLp24(MPR) (C); and VHLp24(R167W) (D).