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. Author manuscript; available in PMC: 2013 Apr 2.
Published in final edited form as: Biochem Biophys Res Commun. 2008 Jul 31;375(1):33–37. doi: 10.1016/j.bbrc.2008.07.113

Figure 2. Identification of a canonical NLS encoded by amino acids 194-243 of Gemin4.

Figure 2

Figure 2

Figure 2

A) Sub-confluent HeLa cells were transiently transfected with 0.5 μg of a panel of truncated Gemin4 constructs: G4ΔN193, G4ΔN243, or G4ΔN293, cloned into the pCI-HA vector. Transfected cells were incubated for 24 hrs and expressed proteins were visualised using an anti-HA mouse monoclonal and an anti-mouse FITC-conjugated secondary antibody. Full-length Gemin4 (FL) is shown as a reference and is taken directly from Fig. 1. Cytoplasmic aggregates are indicated (white arrows). The bar represents ~30μm. B) A schematic of the region containing the predicted NLS. The encoded amino acids are shown, with the putative SV40-like simple (bold font) and the canonical complex (underlined) NLS’s indicated. The corresponding amino acid numbers are shown. C) Expression of aa 194-243 of Gemin4 (G4NLS) is sufficient for nuclear localisation. Removal of the predicted NLS from Gemin4 (G4ΔNLS) results in exclusive localisation of Gemin4 to the cytoplasm. G4NLS and G4ΔNLS are cloned into the pEGFP vector (BD Biosciences). An empty pEGFP vector (GFP), is shown as a reference. DAPI was used to counter-stain nuclei (blue). The bar represents~30μm.