1. GFP
Chlorophyll
Merged A B C D
Supplemental Figure S1. Effects of incubation conditions on the accumulation of vacuole-targeted GFP in vacuolar lumens. A, A Fresh leaf excised from vacuole-targeted GFP expressing plants (Tamura et al., 2003) was observed immediately by LSCM. B, C and D, Excised leaves from vacuole-targeted GFP expressing plants at 30 days after sowing were incubated at 23˚C for 20 h in 10 mM MES-NaOH with 1 μM concanamycin A and 100 μM E-64d in darkness (B), with MS medium containing sucrose in darkness (C) or in the light (D). GFP appears green and chlorophyll fluorescence appears red. In B, C and D, merged images are shown. Bars=50 μm.