Figure 8.

Quantification of Fluorescent PA Bound to RbohD and RbohF Fragments Expressed in GCPs.

(A) RT-PCR analysis of RbohD gene expression in the wild type and rbohD mutant. The experiment was repeated three times under the same conditions.

(B) Quantification of fluorescent PA bound to wild-type and non-PA-binding RbohD fragments. Top panel: immunoblotting of HA-tagged RbohD fragments (160 amino acids) of the wild type and non-PA-binding mutant [R(149,150,156,157)A] expressed in rbohD GCPs. Bottom panel: quantification of fluorescent NBD-PA coimmunoprecipated with RbohD fragments in the absence or presence of 10 μM ABA for 10 min.

(C) Amino acid alignment of the PA binding region in RbohD and corresponding region in RbohF. The double mutation [AL(156,157)RR] in the RbohF sequence is indicated.

(D) Quantification of fluorescent PA bound to wild-type and double mutant RbohF [AL(156,157)RR] fragments. Top panel: immunoblotting of RbohF fragments of the wild type and double mutant [AL(156,157)RR]. Bottom panel: quantification of fluorescent NBD-PA coimmunoprecipated with RbohF fragments in the absence or presence of 10 μM ABA for 10 min.

Data in (B) and (D) are means ± se of three independent experiments.