Figure 4.

Effects of Protein Phosphatase Inhibitors on the Phosphorylation Status of phot1.

The aerial parts of 2-d-old etiolated seedlings were incubated under darkness for 2 h in liquid medium with a protein phosphatase inhibitor (PPase inhibitor), 30 µM CN or 1 µM OKA, and subsequently treated with blue light (BL) at 0.1 µmol m^–2 s^–1 for 0.5 h. Total proteins (15 µg, see [A]; 20 µg, see [B]) extracted from the seedlings were separated on 6% SDS-PAGE gels with 2 µM Phos-tag, followed by immunoblotting with anti-PHOT1 antibodies. The PVDF membranes were stained using a Pierce reversible protein staining kit as a loading control.

(A) Effects of protein phosphatase inhibitors (PPase inhibitors) on the aerial parts of the wild-type Col seedlings and rpt2 mutants. Red light pretreatments were conducted to induce RPT2 expression before exposure to the PPase inhibitor.

(B) Effects of protein phosphatase inhibitors (PPase inhibitors) on the aerial parts of the wild-type Col seedlings and the 35Spro:RPT2 lines (OX).