Skip to main content
. 2020 Mar 24;11(3):345. doi: 10.3390/genes11030345

Figure 2.

Figure 2

In vitro kinase activity assay of MLK3. The kinase activity assay of the recombinant protein MLK3-MBP (a) and MLK3 (K146R)-MBP (b) by dot blotting. MLK3 and MLK3 (K146R) were fused with maltose-binding protein (MBP) separately. H3, an unmodified histone H3 peptide (residues 1-21), biotin conjugate (Upstate 12-403) was used as a substrate. The reaction mixture (1 µl) was spotted on a nitrocellulose membrane as indicated by Ponceau S staining and phosphorylation was examined by immunoblotting with an antibody against H3T3ph (Upstate, 07–424). The reaction mixture without the recombinant protein(s) served as the negative control (NC) and peptide H3T3ph, phospho-histone H3 (Thr3) peptide (residues 1–21), biotin conjugate was used as the positive control (PC).