Fig. 2. Transgenic Arabidopsis expressing the  APXII-Fluc  chimera gene. a) Schematic representation of the  APXII-Fluc  chimera gene. The chimera gene, in which the donor site of intron 11 to the acceptor site of intron 12 including SRE of the spinach  APXII  gene is fused with the Fluc cDNA at its intron 3-spliced site, driven by the CaMV 35S promoter was transformed into Arabidopsis using  A. tumefaciens . Exon regions are shown as boxes and introns as lines. The open reading frame and untranslated regions are indicated by black and white boxes, respectively. Functional and premature stop codons are indicated by asterisks. The position of the SRE sequence is indicated by a bold line. Arrows (F1–F3) indicate the location of primers used in this study ( Supplementary Table 1 ). LB, left tDNA border; RB, right tDNA border. b) Results of semiquantitative RT-PCR analysis to detect the AS variants (sAPX-II, -III, and tAPX-I types) in the  APXII-Fluc  chimera gene-transformed Arabidopsis ( APXII - Fluc ). Semiquantitative RT-PCR analysis (26 cycles) was carried out using sets of primers in the extract from the rosette leaves of 2 weeks old  APXII - Fluc  plants (T3 generation). Amplified products were electrophoresed on a 2% (w/v) agarose gel. M, molecular marker. c) Results of immunoblot analysis to detect the Fluc protein in the  APXII - Fluc  and WT Arabidopsis. Immunoblot analysis was performed using the antibody raised against the Fluc protein produced from the tAPX-I-type mRNA in extracts from the rosette leaves of 2-week-old  APXII - Fluc  and WT Arabidopsis. d) Luminescence image of  APXII - Fluc  and WT Arabidopsis. Fluc-dependent luminescence was imaged in 2-week-old plants by EM-CCD camera. The experiments were repeated at least 3 times using plants grown independently, and the representative results are shown.