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. 2006 Apr 10;103(16):6398–6403. doi: 10.1073/pnas.0601620103

Fig. 4.

Fig. 4.

Cellular localization of SFT, organ distribution of SFT-interacting proteins, and SFT transcript assay in receptor graft tissues. (A) Expression profile of a GUS-tagged SFT promoter. Shown are a SAP (Left), the youngest leaf with detected expression (Center), and a series of developing leaves (Right). (Scale bars: 1 mm.) (B) Diurnal RNA expression of SFT-interacting proteins in 3- to 4-cm leaves and SAPs of tomato. (C) Correlation between SFT-MYC antigen (Upper) and SFT-MYC RNA (Lower) in progenies of a 35S:SFT-MYC/+ Samsun plant segregating for flowering time. (D–F) The SFT-MYC antigen is localized primarily in the nucleus. (D) Intracellular localization of SFT-MYC antigen in a sepal of 35S:SFT-MYC tobacco (rhodamine staining). (E and F) An optical confocal section and Nomarski image (alkaline phosphatase), respectively, of tobacco cells expressing the SFT-MYC antigen. The arrow points to the spindle of a decorated dividing cell. (G) Calibration of RT-PCR detection levels of 35S:SFT-born transcripts (Ω and RI primers) in RNA from leaves of a 35S:SFT donor and a rescued sft-k receptor. Lanes 1–6, 35S:SFT donor RNA template in 1/5 dilution series starting with 5 μg of RNA; lanes 7 and 8, 5 μg of template RNA from WT and rescued sft receptor SAPs, respectively. (H) RT-PCR detection of endogenous SFT RNA in rescued receptor sft organs (5 μg of RNA; FII and RII primers). Lanes 1–4, template RNA of sft-k receptor from young leaves, SAPs, stem, and flowers, respectively; lanes 5 and 6, WT and donor 35S:SFT leaves, respectively; lane 7, no template control. (I) Nested RT-PCR detection (primers FII and RII) of 35S:SFT transcripts (odd lanes, first-round primers Ω and RI) in comparison with SFT transcripts (even lanes, first-round primers FI and RI) in the four rescued sft recipient organs as above (lanes 1–8), in WT (lanes 9 and 10), and in 35S:SFT donor (lanes 11 and 12). Dilution ratios of first-round PCR were 1/1,000 for the 35S:SFT and 1/105 for the endogenous SFT transcripts. (J) A scheme of the 35S:SFT transgene showing the primers used in the PCR experiments.