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. 2014 Aug 5;13(9):4074–4090. doi: 10.1021/pr500437x

Figure 7.

Figure 7

In vitro kinase activity assay of full-length NEK7 (6×His-NEK7), NEK7 kinase domain (6×His-NEK7Δ(1–44)), chimeras N6C7 (6×His-N6C7) and N7C6 (6×His-N7C6), full-length NEK6 (6×His-NEK6), and NEK6 kinase domain (NEK6Δ(1–33)). The enzymes were incubated at a concentration of 80 nM with 50 nM ULight-p70 S6K Peptide, and the phosphorylation was scored in the presence of 100 μM ATP. Kinase reactions were terminated after 60 min by the addition of EDTA. A reaction without addition of ULight-p70 S6K Peptide was used as a control (Control). Data represent the mean and standard deviation of three independent experiments. The signal was measured on a 2104 EnVision Multilabel Microplate Reader. Excitation wavelength was set to 320 nm, and fluorescence emission was recorded at 665 nm.