Figure 1.

Half-life of DHHC2 mRNA. (A) The abundance of DHHC2 mRNA after siRNA-mediated DHHC2 knockdown was measured by qRT-PCR at 0, 12, 24, 48, 72, and 96 h after siRNA oligo transfection. These experiments were run in triplicate. The data were approximated most accurately using a nonlinear, first-order exponential decay. The half-life was calculated to be 13.87 h (R² = 0.998, n = 3; error bars, SE). (B) The relative abundance of DHHC2 mRNA in siRNA-transfected cells versus mock-transfected control cells was determined after 96 h by qRT-PCR. DHHC2 expression was reduced ∼84% with siRNA knockdown. (C) CKAP4 interacts with CFP-DHHC2. CFP-tagged DHHC2 was immunoprecipitated from transiently-transfected HeLa cells using an anti-GFP mAb (see Materials and Methods). Western blot analysis with an anti-CKAP4 antibody detected a robust band at ∼63 kDa, characteristic of CKAP4 in lysates from cells transfected with CFP-DHHC2 but not in lysates from mock-transfected cells. Exposure time, 15 s.