FIGURE 5.

Expression of NTT and HEC1 versus Kunity-PI;1 transcripts in response to phytohormones. (A) Semi-quantitative RT-PCR analysis of NTT, HEC1, and Kunitz-PI;1 expression in 3-days old etiolated seedlings grown on ACC-containing, silver nitrate-containing or IAA-containing Murashige–Skoog medium. For comparison, actin transcript levels were assessed as internal standard. (B) Kunitz-PI;1 protein levels in 3-days old etiolated seedlings after growth on ACC-containing or Ag⁺-containing medium analyzed by Western blotting. (C) Kunitz-PI;1 protein accumulation in etiolated seedlings of WT, ntt and hec1 mutant, as well as 35S::hec1 overexpressor. For SDS-PAGE (B,C), 40 μg protein was loaded per lane and subjected to Western blotting using Kunitz-PI;1-specific antibodies (upper panels); loading was confirmed by Ponceau-staining of the nitrocellulose-blotted proteins (lower panels). Note that the lower panels in (C) are composite diagrams.