Figure 5

Differential expression of Zm-miR164 between 87-1 and Zong3. A. miR164 cleavage sites in ZmNAC1 mRNA were determined by RNA ligase-mediated 5^′ RACE. The resulting agarose gel showed the nested PCR products that we cloned and sequenced, and the product was expected to be approximately 300 bp. B. The frequency of 5’ RACE clones corresponding to the cleavage site (indicated by arrows) is shown as a fraction, with the number of clones matching the target message in the denominator. C. RNA gel blot analysis of Zm-miR164 in 10 μg of low-molecular-weight RNA that was prepared from 8-day-old root samples from two separate inbred lines. A 5S rRNA sample was used as a loading control (bottom gels). D. Expression analysis of miR164b and miR164d with real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Eight-day-old root seedling samples were used for the RNA extraction.