Northern Blot Analysis of GFP mRNA

RNA (‘northern’) blot analysis of GFP mRNA was performed as described previously (Du et al., 2014a (link)). Total RNA was extracted from agroinfiltrated leaves using TRIzol reagent (Invitrogen) according to the manufacturer’s instruction. Five micrograms of each total RNA sample was separated on 1.5% agarose gel containing 7% formaldehyde, and transferred onto Hybond+nylon membrane (GE). GFP mRNA was detected by the digoxigenin (DIG)-labeled DNA probes that were prepared using the DIG high primer DNA labeling and detection starter kit II (Roche) as recommended by the manufacturer, and cleaned up using G25 Sephadex columns (GE). DIG-labeled probes were detected using a chemiluminescence-based DIG detection kit (Roche) according to the manufacturer’s instructions.