FIG. 5.

Lack of Ptc1 affects Npr1 levels and phosphorylation state. Wild-type BY4741 (WT) and its ptc1 (MAR143) or sit4 derivative were transformed with plasmid pEJ23, which carries an N-terminally HA-tagged version of Npr1. Cells were incubated for 30 min with 100 ng/ml rapamycin or vehicle, and extracts were prepared. Thirty micrograms of protein [90 μg in the case of ptc1 cells, denoted ptc1(3x)] was electrophoresed, transferred, and incubated with anti-HA monoclonal antibodies. Faster-migrating bands correspond to the dephosphorylated forms of Npr1.