Figure 2.

PKS5 and J3 Have Overlapping Tissue-Specific Expression and Subcellular Localization.

(A) to (J) Expression of J3 and PKS5 in 3-d-old seedlings ([A] and [F]), 10-d-old seedlings ([B] and [G]), rosette leaves ([C] and [H]), roots ([D] and [I]), and cross sections of roots ([E] and [J]).

(A) to (E) PKS5 promoter-GUS expression in wild-type seedlings.

(F) to (J) J3 promoter-GUS expression in wild-type seedlings.

(K) to (P) Subcellular localization of GFP-J3 ([K] to [M]) and PKS5-YFP ([N] to [P]) in the upper portion of the root. Confocal GFP images ([K] and [N]), bright-field images ([L] and [O]), and combined GFP and bright-field images ([M] and [P]).

(Q) Both of PKS5 and J3 were detected in soluble and plasma membrane–enriched fractions. Isolation of plasma membrane vesicles was by two-phase partitioning from seedlings of j3-1 plants expressing 35SP:3×flag-J3 or pks5-1 plants expressing DexP:3×flag-PKS5. Equal amount of soluble (S) and plasma membrane (P) proteins were separated by SDS-PAGE followed by analysis with anti-flag, anti-MAPK3, or anti-PM H⁺-ATPase antibodies.

(R) Both of PKS5 and J3 were detected in nucleus. Nuclei were isolated from seedlings of j3-1 plants expressing 35SP:3×flag-J3 or pks5-1 plants expressing DexP:3×flag-PKS5. The nuclei pellet was then washed three times. Equal amount of three washing (S1-S3) and nuclei (N) proteins were separated by SDS-PAGE followed by analysis with anti-flag or anti-Histone H3 antibodies.