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. 2008 Nov 19;105(48):18788–18793. doi: 10.1073/pnas.0810104105

Fig. 5.

Fig. 5.

Effect nocodazole and siRNA on the intracellular localization of BIG1 and KIF21A. (A) HEK293 cells transfected 24 h earlier with GFP-KIF21A were incubated with DMSO vehicle (No) or nocodazole (Noc, 10 μg/ml) for 1 h before reaction with anti-BIG1 and anti α-tubulin antibodies. (Scale bar, 16 μm.) (B) Cells grown in two-well poly D-lysine-coated slides were transfected with GFP-KIF21A 24 h before incubation for 48 h with nontargeting (NT) or BIG1-specific siRNA, followed by fixation and immunostaining with α-tubulin antibodies (Scale bar, 16 μm). (C) Proteins in cell extracts Total (postnuclear), cytosol, and membrane fractions prepared 72 h after addition of nontargeting (lane 1) or BIG1-specific siRNA (lane 2) treatment, were separated in 4–12% NuPAGE gels and immunoblotted with indicated antibodies. (D) After incubation for 72 h with addition of nontargeting (NT) or KIF21A siRNA, cells were fixed and immunostained with anti-BIG1, giantin and α-tubulin antibodies. (Scale bar, 16 μm.) (E) Samples of cell extracts total (postnuclear), cytosol, and membrane fractions prepared 72 h after nontargeting (lane 1) or KIF21A siRNA (lane 2) treatment were separated in 4–12% NuPAGE gels, and immunoblotted with indicated antibodies.