Kanegae et al. 10.1073/pnas.0603569103. |
Fig. 4. Relative quantitative RT-PCR analysis of PHY3 transgene mRNA levels.
Total RNA was extracted from green leaves of 4-week-old seedlings by RNeasy Plant Mini Kit (Qiagen) and treated with DNA-free (Ambion) to remove contaminating genomic DNA. For relative quantitative RT-PCR, multiplex RT-PCR with an endogenous standard (Quantum RNA 18S Internal Standards; Ambion) was performed according to the manufacturer's protocol. PHY3 gene-specific primers used were: 5'-ATCTCATTAGTGCATTCCAGCATA-3' and 5'-GGAATATCCTTCTCCTTCTCCATC-3'. Reverse-transcription and PCR amplification was done by using SuperScriptII reverse transcriptase with random hexamers as a primer (Invitrogen) and by Platinum Taq DNA polymerase (Invitrogen), respectively.
Fig. 5. Red light does not affect the degree of hypocotyl curvature induced by blue light in C303S transgenic Arabidopsis.
C303S expressing phot1-5 phot2-1 transgenic Arabidopsis (C303S-1) was grown in the dark for 3 days, and then light irradiation was done as follows: RL, continuous unilateral red light (0.1 μmol·m-2·s-1) for 16 h; BL, continuous unilateral blue light (2.0 μmol·m-2·s-1) for 16 h; RL+BL, RL (0.1 μmol·m-2·s-1) and BL (2.0 μmol·m-2·s-1) were irradiated simultaneously for 16 h. At the end of the lateral light irradiation, phototropic curvatures were measured. Each bar indicates a mean of 15 measurements with standard deviations.