Figure 5.

Anti-LeMPK antibodies specifically recognize LeMPKs. Polyclonal antibodies were raised against the N-termini of LeMPKs shown in Figure 4. Recombinant LeMPKs were expressed in E. coli and affinity purified. Five hundred nanograms of each recombinant LeMPK was analyzed by immunoblotting on three separate membranes with anti-LeMPK1, anti-LeMPK2, and anti-LeMPK3 antisera. Antibody-LeMPK immunocomplexes were visualized using an alkaline phosphatase-coupled colorimetric assay. On a separate gel, recombinant LeMPKs were stained with Coomassie Brilliant Blue. Arrowheads with numbers indicate the apparent molecular masses of the recombinant proteins.