Figure 5.

Role of farnesylation in localization and activity of Kuk constructs and LaminDm0ΔN in cultured cells. (A–D) NIH-3T3 cells transiently transfected with different constructs. (A′–D′) Transiently transfected NIH-3T3 cells, treated with the FTI ABT-100 for 24 h. Lamin A/C staining (green) marks the NM (A–D′). Localization pattern of the transfected constructs is shown in red (Kuk in A–C′ and HA in D and D′). Bar, 10 μm. (E) S2 cells treated with the FTI ABT-100 for 5 d. Lamin Dm0 staining (red) marks the NM and Kuk is shown in green. Bar, 5 μm. (F) FTI treatment increases the electrophoretic mobility of Kuk. In total extract of nontreated S2 cells a band of ∼120 kDa, corresponding to farnesylated Kuk is detected. In the total extract of FTI-treated cells, the intensity of the ∼120-kDa band is reduced and an ∼110-kDa band representing not farnesylated Kuk is also detected. Kuk runs at a significantly higher MW than predicted (Brandt et al., 2006).