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. 2008 Nov 12;36(22):7230–7239. doi: 10.1093/nar/gkn896

Figure 5.

Figure 5.

Effect of depurination location on BMV RNA replication in vivo. (A) Barley protoplasts were transfected with PAP-treated, ligated RNA3 templates described in Figure 4A, plus intact BMV RNA1 and 2 in vitro transcripts and allowed to replicate. Total protoplast RNA was analyzed by northern blot for the presence of positive-strand BMV RNAs. Std is in vitro transcript of positive-strand RNAs 1, 2 and 3 to serve as size markers. Values are means of intensities for RNAs 1, 2 and 3 ± SE for three separate experiments, quantified with a phosphorimager. Blots were also probed for 28S rRNA as a loading control for total RNA. (B) Stability of type I template in barley protoplasts. Type I template (ligated) or non-ligated RNA3 (control) was transfected into barley protoplasts and aliquots of cells were removed at the indicated time points. Total protoplast RNA was probed by northern blot for positive-strand BMV RNA3. The blot was also probed for 28S rRNA as a loading control for total RNA.