Figure 2

A) Specific NFI DNA-binding activity of recombinant CeNFI-H6. Partially purified recombinant H6-tagged CeNFI (containing parts of exons 2–8) and human NFI-C220 were incubated with a duplex oligonucleotide containing an NFI binding site (2.6, even lanes) or an oligo with a single point mutation that abolishes NFI binding (C2, odd lanes) and analyzed on a 6.5% non-denaturing polyacrylamide gel. Lanes 1 and 2, crude E. coli extract (neg. control); lanes 3 and 4, ~5 ng partially purified CeNFI-H6; lanes 5 and 6, ~40 ng partially purified CeNFI-H6; lanes 7 and 8, ~5 ng purified human NFI-C220H6. See bottom of panel for sequences of oligonucleotides. B) Specific NFI DNA-binding activity in worm extracts. Nuclear extracts of a mixed population of C. elegans were prepared and used in a gel mobility shift assay with an oligonucleotide that contains an NFI-binding site (lanes 1 & 2, 2.6) or the same oligo with a single point mutation that abolishes NFI binding (lane 3, C2). Lane 1, no extract; Lanes 2 and 3, C. elegans extract (~10 μg). See A for sequences of oligonucleotides.