Fig. 2. Confirmation of MPSS data using different techniques. (A) Induction or repression of seven genes was verified by RNA gel blot analyses. Total RNAs from an independent ABA (50 µM) treatment were isolated from WT plants after induction for 3 hours and 5 hours (+). Control plants were treated in parallel (-). The RNA gel blot was probed with DNA fragments of the indicated genes. (B) RT-PCR was used to confirm regulation of genes with low transcript abundance. 200-500 ng total RNA of samples from an independent 4 hours ABA (50 µM) treatment was used for each of the indicated RT-PCR reactions. The 25S RNA band stained with ethidium bromide served as a loading control. The names of genes shown in A and B are: KIN2 (At5g15970), putative NPK1-related MAP kinase (At1g05100), putative protein phosphatase 2C (At1g07430), ethylene responsive element binding factor 4 (At3g15210), citrate synthase-like protein (At3g58750), putative RNA-binding protein (At1g09340), MPK6 (At2g43790), ZFP2 (At5g57520), putative bHLH transcription factor (At2g46510), putative homeodomain transcription factor (At2g35940), potassium channel-like protein (At4g18160), and AAO2 (At3g43600).