Figure 3. Constructs of various LIN54 mutants. (A) Schematic representation of LIN54 mutants. Fragments of LIN54 were ligated in frame with one of various epitope tags (EGFP, FLAG and HA). EGFP and HA epitope tag were fused to the C terminus and FLAG epitope tag was fused to the N terminus. DNA constructs encoding these mutants were inserted into adenoviral vectors. (B) GC-1 cells were infected with each recombinant adenovirus and then LIN54 and mutant proteins were detected by western blot with indicated antibodies. β-tubulin is a loading control. (C) Alignment of the amino acid sequence of CXC domains with those of other CHC family proteins. Conserved cysteine residues are shaded in black. The asterisks represent the point-mutated cysteine residues (C525 and C611).