Figure 2

Transient expression of truncated PGL34-GFP fusions in protoplasts. A. DNA constructs encoding fragments of or full length AtPGL34 coding sequence fused to GFP. The transit peptide of PGL34 is shaded. The Kyte and Doolittle hydropathy plot of PGL34 is shown and domains with higher hydropathy scores (H1–H3), as well as a domain conserved among PAP-fibrillin proteins (black bar), are indicated. B. Fluorescence of the GFP fusion proteins (GFP) was detected in transformed protoplasts by confocal laser scanning microscopy. Arrows indicate strong GFP signals overlapping with weak chlorophyll autofluorescence signals (chlorophyll). Merge: overlap of chlorophyll and GFP signals. Scale bars: 5 μm. C. Detection of GFP fusion proteins in transformed protoplasts by immunoblotting using anti-GFP antibodies. D. Cotransformation of protoplasts with full length and truncated PGL34. Protoplasts coexpressing PGL34-CFP and PGL34_1–133-YFP, PGL34_1–170-YFP or PGL34_1–290-YFP were analysed by confocal microscopy. CFP fluorescence (CFP) and YFP fluorescence (YFP) were monitored sequentially using distinct excitation wavelengths and detection windows. Chlorophyll: chlorophyll autofluorescence, merge: superposition of YFP and CFP signals (green and red pseudocolours, respectively). Bar length: 5 μm.