Figure 2
From: Identification and characterization of plant Haspin kinase as a histone H3 threonine kinase
GST-AtHaspin phosphorylates histone H3 at Thr3 and Thr11 in vitro. (A) GST-AtHaspin and GST-AtHaspin-KD were incubated with or without ATP, and phosphorylated proteins were stained with ProQ Diamond Phosphoprotein stain. (B) GST-AtHaspin and GST-AtAUR3 were incubated with GST-H3 tail (left and right lanes). Negative control: GST-H3 tail only (middle lane). Phosphorylated GST-H3 tail was immunostained using anti-H3T3ph, H3T11ph, H3S10ph, and H3S28ph antibodies. (C) GST-AtHaspin and GST-AtHaspin-KD were incubated with GST-H3 tails or mutants as substrates. Phosphorylated GST-H3 tails were immunostained with anti-H3T3ph and anti-H3T11ph antibodies.