Fig. 2
From: Screening and identification of emodin as an EBV DNase inhibitor to prevent its biological functions
The effects of pH, temperature, divalent cations, and ion strength on EBV DNase activity. The biochemical characteristics of the purified EBV DNase were assessed using the fluorescence-based method. a To determine the optimal pH, proteins were diluted in different pH conditions and were incubated with salmon sperm DNA for 1 h at 37 °Ϲ. PicoGreen dye was then added and fluorescence was detected using a microplate reader. Solid squares (EBV DNase) represent EBV DNase groups and open circles (CTL) represent mock controls without enzymes, respectively. b To determine the optimum temperature, the protocol was the same as described above, except for incubation at different temperatures. c To determine the optimal concentration of divalent cations for EBV DNase, nuclease activity was assessed following incubation of DNase protein with salmon sperm DNA with various concentrations of magnesium (left panel) and manganese (right panel). Solid squares (EBV DNase) represent EBV DNase groups and open circles (CTL) represent mock controls without enzymes, respectively. d To identify the inhibition of nuclease activity of EBV DNase at different ionic strengths, DNase protein was incubated with salmon sperm DNA at various concentrations of KCl (left panel), NaCl (middle panel), and (NH4)2SO4 (right panel). PicoGreen dye was then added, and fluorescence was detected (excitation: 480 nm, emission: 520 nm). Data are presented as the means and standard deviations of three independent experiments