Fig. 4. Transient expression of hARL5 and its mutants in COS-7 cells. (A) Diagrams of wild-type (WT) hARL5 and four hARL5 mutants with amino acid numbers below. Residues 188 to 201 contain the bipartite NLS of ARL5. ARL(dC) lacks 24 amino acids (177 to 201) at the C-terminus. Positions of the mutations Q80L and T35N, and MIR deletion (128-133) are also indicated. (B) COS-7 cells transfected with GFP fusion constructs of hARL5(WT), hARL5(Q80L), hARL5(T35N), or hARL5(dC) were grown on glass coverslips, fixed with formaldehyde, incubated with mouse anti-nucleolin (nucleolar marker), and detected with Alexa-488-conjugated anti-mouse IgG antibody. Coverslips were mounted on Mowiol (supplemented with Hoescht 33258) and inspected with a Zeiss Axiophot equipped for epifluorescence. Phase-contrast microscopy and GFP-hARL5 fluorescence (green) are shown. In the bottom two rows, arrows indicate nucleolins, and arrowheads indicate Golgi marker p58. Bar, 10 µm

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