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. 2013 Oct 29;23(12):1369–1379. doi: 10.1038/cr.2013.143

Figure 2.

Figure 2

PYL13 specifically binds to and inhibits PP2CA independent of ABA. (A) Size exclusion chromatography (SEC) analyses of the complexes between PYL13 and ABI1, HAB1, HAB2, and PP2CA. The indicated fractions of SEC eluents were applied to SDS-PAGE followed by coomassie-blue staining. The peak position of the PYL13-PP2CA complex is indicated by the magenta line. (B) PYL13 specifically inhibits PP2CA independent of ABA. The details of the experiments are described in Materials and Methods. The readout of the reactions without phosphatase was set as the baseline. The phosphatase activity of the indicated PYL13-PP2C complex is normalized against that of the corresponding PP2C at the same molar concentration. The concentrations of all the PYL13-PP2C complexes are adjusted to 0.2 μM for assays. ABA was added with a final concentration of 2 μM. All the experiments are independently performed for at least three times. The error bars represent SD. (C) PP2CA is specifically inhibited by PYL13 in the absence of ABA. PYL2 and PYL10 rely on ABA to inhibit PP2CA. Notably, PYL10 is a potent ABA-independent inhibitor of ABI1, HAB1, and HAB2, but not PP2CA13.