Figure 2
Comparison of transfection efficiencies between rice green tissue protoplasts and etiolated protoplasts. Transient expression of different constructs in rice protoplasts. Samples were visualized under a confocal microscope. A-B, The 35S::GFP construct pUC-GFP was transiently expressed in protoplasts derived from 8-day-old rice green seedlings (A) or 8-day-old etiolated rice seedlings (B). Individual and merged images of GFP and chlorophyll autofluorescence (Chl) as well as bright field images of protoplasts are shown. Scale bars = 10 μm. C, The transfection efficiency of rice green tissue protoplasts was comparable to that of etiolated protoplasts. The ratio of GFP-positive cells to the total number of protoplasts (n ≥ 100) was scored as the transfection efficiency. Values are means, with standard errors indicated by bars, representing at least 5 replicates. D, Detection of transiently expressed bZIP63-c-myc-YFPN (lane 1) and c-myc-YFPN (lane 2) by Western blot. Lane 3 was a negative control. The upper panel shows an immunoblot using a monoclonal mouse anti-c-myc antibody; the lower panel shows a Coomassie Brilliant Blue (CBB)-stained PVDF membrane after immunoblotting as a loading control.