Figure 3.

Autophosphorylation of NTHK2-KD. GST fusion protein NTHK2-KD (0.5 μg) was incubated with [γ-³²P]ATP in the presence of 5 mm Mg²⁺, 5 mm Ca²⁺, 5 mm Mn²⁺, or 10 mm EDTA. GST (4 μg) was included as a control. The phosphorylated proteins were then separated on SDS-PAGE, transferred onto PVDF membrane, and autoradiographed (top) or Coomassie blue stained (bottom).