Skip to main content
. 2019 Mar 22;14(3):e0213598. doi: 10.1371/journal.pone.0213598

Fig 1. Characterization of the phosphatase activity acting on EECs.

Fig 1

A, diagram of steps performed to analyze phosphatase activity (left) and EC-EMSA of EECs incubated for 3 or 30 minutes after stopping the pulse with EDTA followed by the indicated washes. LSW and HSW are low (60 mM KCl) or high (1.6 M KCl) salt washes. Details are in Materials and Methods. B, the effect of magnesium on phosphatase activity (left) and the contribution of FCP1 (right). EECs were washed as indicated and incubated with EDTA, Mg, HeLa nuclear extract (HNE), FCP1, or TFIIF (IIF) for the indicated times. Phosphorylated EECs (P); unphosphorylated EECs (UnP). C, Indicated Mono Q fractions of HNE were incubated for 30 minutes with high salt washed EECs that were resuspended in low salt wash (Phosphatase assay). The same fractions were blotted with antibodies to Ssu72 and shown at two different exposures (Western blot).