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. 2008 Mar 12;3(3):e1755. doi: 10.1371/journal.pone.0001755

Figure 2. DCL2 and DCL4 have Distinct Roles in Self-silencing of the ΔGUS-SUG Hairpin Transgene.

Figure 2

(A) The diagram shows the coding region of the ΔGUS-SUG hairpin (hp) transgene drawn to illustrate the double-stranded configuration of the transcript. The locations of the hybridization probes and the 231-bp deletion are indicated. Probes 1 and 3 are as specified in Figure 1A. Accumulation of GUS mRNA in the ΔGUS-SUG line 306-1 was determined using RNA gel blot analysis and RNA probes. Duplicate samples of two different RNA preparations were run on one gel and blotted onto one membrane, after which the membrane was cut in half. One half was hybridized with probe 1 and the other with probe 3*; probe 1* gave the same result as probe 1 (data not shown). Ethidium bromide stained rRNA is shown as a loading control. (B) Accumulation of GUS mRNA and siRNAs in wild type (wt) and mutant lines carrying the ΔGUS-SUG transgene of line 306-1 was determined using RNA gel blot analysis with RNA probes. Probe 3* was used to detect GUS mRNA, while probes for GUS siRNAs all had sense polarity. Grouped lanes are all from the same gel, blot, and exposure. LMW RNA blots were successively stripped and hybridized with the indicated probes. The positions of 21- to 24-nt RNA size markers are indicated. Ethidium bromide stained rRNA and the major RNA species in LMW RNA are shown as loading controls.