FIGURE 5.

JH III regulated BrZ7 phosphorylation through a GPCR-, PLC-, and PKC-triggered pathway. A, the inhibitors of GPCR, PLC, and PKC suppressed JH III-induced BrZ7 phosphorylation. Cells were transfected with pIEx-4-BrZ7-His for 48 h, pretreated with different inhibitors for 1 h, and incubated with 1 μm JH III for 3 h. Control cells were transfected with pIEx-4-RFP-His. Western blot was performed on 7.5% SDS-PAGE gel with anti-His antibody. B, predicted putative phosphorylation sites of BrZ7 are underlined. C, mutant plasmids were constructed. Boxed amino acids correspond to the mutated amino acids. D, the phosphorylation of BrZ7 was not affected by the overexpression of different mutant plasmids using pIEx-4-His in HaEpi cells as revealed by Western blot.