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. 2017 Jan 6;29(1):144–155. doi: 10.1105/tpc.16.00720

Figure 3.

Figure 3.

The C-Terminal Part of KNL2 Binds Centromeric and Noncentromeric DNA Sequences in Vitro.

(A) Nonradioactive EMSA with 4 pmol of the centromeric repeat pAL1 (all lanes) and 4 to 32 pmol of the recombinant C-terminal part of KNL2 (lanes 2–5) or 16 and 32 pmol of the recombinant N-terminal part of KNL2 (lanes 6 and 7), respectively, showed that the C terminus (lanes 4 and 5), but not the N terminus, of KNL2 interacts with the centromeric repeat pAL1 (n = 3).

(B) Nonradioactive EMSA with 4 pmol of pAL1 (all lanes) and 16 pmol of the C-terminal part of KNL2 (lanes 2–9). The unspecific competitor poly dI/dC was added in increasing concentrations of 0.1, 0.5, 1, 5, 10, 50, and 100 ng/μL (lanes 3–9). At an amount of 50 pmol poly dI/dC, the DNA binding of the C-terminal part of KNL2 and pAL1 get competed out (n = 2).

(C) Nonradioactive EMSA with 4 pmol of centromeric repeat pAL1 (lanes 1 and 2), transposable element Athila (lanes 3 and 4), telomeric repeat (lanes 5 and 6), euchromatic TUA4 sequence (lanes 7 and 8), and 16 pmol of the C-terminal part of KNL2 (even numbered lanes) shows that the C-terminal part of KNL2 interacts with centromeric and noncentromeric DNA (n = 3).