Skip to main content
. 2017 Jan 30;29(2):260–276. doi: 10.1105/tpc.16.00719

Figure 9.

Figure 9.

WER Forms a Stable Complex with Its Target DNA When NRP1 Associates with H2A/H2B Dimer.

(A) SEC profiles (Superdex 200 10/300 GL) of WER (A280 in cyan) and its DNA substrate (A254 in black), as well as their complex at 1:1 molar ratio (A254 in red; A280 in blue).

(B) EMSA of NRP1, H2A/H2B dimer, WER, and its substrate DNA. Lanes 1 to 14, 2 μM DNA (black arrow, set as relative molar ratio at 1). Lanes 2 to 4, WER titrated against DNA at molar ratios of 2:1, 4:1, and 6:1. Lanes 5 to 9, H2A/H2B dimer titrated against DNA at molar ratios of 1:1, 2:1, 4:1, 5:1, and 6:1, while amount of WER was held at 6:1 molar ratio. Lanes 10 to 14, NRP1 titrated against DNA at molar ratios of 1:1, 2:1, 4:1, 5:1, and 6:1, while WER and H2A/H2B dimer were both held at molar ratio of 6:1 against DNA. Lane 15, H2A/H2B dimer and NRP1 as in lane 14 without WER and its substrate DNA. All samples were separated on 6% native PAGE gel, stained with GelRed, and visualized by UV (upper panel). The GelRed-stained gel shows bands of DNA bound to H2A/H2B (black arrowhead) and DNA bound to WER (open arrowhead). Gel was further stained by CBB to show bands of NRP12-(H2A/H2B)4 (green arrowhead), NRP12-(H2A/H2B)2 (red arrowhead), and WER bound to DNA (purple arrowhead).