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. 2003 Jul 28;100(16):9632–9636. doi: 10.1073/pnas.1733874100

Fig. 2.

Fig. 2.

Effect of siRNA on EGFP and DsRed2 expression in BY2 tobacco protoplasts. Both the reporter plasmids were driven by the CaMV35S promoter. (A) Schematic representation of the EGFP-coding region and the siRNA sequence (nucleotides 124–144) designed to target EGFP. The slashed area represents the sequence used for the probe in D. NOS-T, nopaline synthase terminator. (B) Schematic representation of DsRed2-coding region and the corresponding siRNA sequence (nucleotides 191–211) to target DsRed2. (C) Protoplasts were transfected either with the EGFP and DsRed2 plasmid DNAs together or in combination with either siGFP or siRNA cognate to red fluorescent protein. Triplicate electroporation experiments were performed in every case. Columns and bars indicate mean and SD values, respectively. GFP and DsRed fluorescence are normalized with nontransfected control cells. (D) RNA gel blot showing siRNA accumulation in protoplasts. Lane 1, protoplasts transfected with pCaMV35S-GFP plasmid DNA with siGFP; lane 2, protoplasts transfected with pCaMV35S-GFP plasmid DNA; lane M, 21-, 30-, and 43-nt end-labeled oligos are shown as molecular size markers.