Figure 1.
Identification of native VHL gene products. Total cellular extracts from 786-0 cells stably expressing VHLp24(MPR) (lane 1), VHLp18(MEA) (lane 2), or control 786-0 cells (lane 3) or from 293 cells (lanes 4–8) were immunoprecipitated (IP) with either VHL mAb 12.21 (lanes 1–3, 5–8), or a nonspecific (unrelated) mAb, 12CA5 (NS, lane 4). Immunoprecipitates were separated by electrophoresis on an SDS/12.5% polyacrylamide gel and transferred to a poly(vinylidene difluoride) (PVDF) membrane. The membrane was cut and Western blotting was performed on the pieces by using rabbit polyclonal antisera to peptides in VHL exon 1 (lanes 1–5), exon 2 (lane 6), exon 3 (lane 7), or preimmune rabbit sera (lane 8). VHLp24(MPR) and VHLp18(MEA) are indicated to the left of the blot. Cross-reacting IgG (light chain) is indicated to the right of the blot. NS mAb (lane 4) is subtype IgG2b and did not cross-react with the anti-rabbit IgG secondary antibody.