Fig. 3.

Characterization of binding mode of Neurospora TTAGGG-binding proteins. a Western blots from pull-downs of recombinantly expressed Neurospora proteins incubated with concatemerized oligonucleotides of telomeric repeats (TTAGGG) or a shuffled control sequence (TGTGAG) to check for direct binding. b Comparative binding test for double-stranded versus single-stranded telomeric DNA. Direct TTAGGG-binding proteins from (a) were incubated with increasing amounts of single-stranded TTAGGG repeats. Only NCU2404 binding is decreased at higher ssTTAGGG oligonucleotide concentrations revealing the other 5 proteins as double-strand binders. c Western blot of the competition assay of NCU02404 shows that its binding can only be competed with ssTTAGGG but not with an unrelated control sequence