Fig. 3

Western blot analysis of GFP in MDMV OH5 derived constructs. Samples of the newest fully emerged leaf from top of the plants were collected at 7, 14, and 21 days post rub-inoculation and analyzed for GFP protein using Anti-GFP antibody (Abcam, USA). Constructs were as diagrammed in Fig. 1, and pWX70 is similar to pWX56 with a similar-sized insert at P1/HCPro without further data reported. GFP from MDMV OH5 constructs with additional 6xHis and protease cleavage sequences with predicted molecular weight of 29.4 kDa were detected at slightly slower migration (32 kDa estimated based on ladder) compared to the positive control GFP (27 kDa estimated based on ladder) with predicted amino acid molecular weight of 26.8 kDa. Twenty-five μg total protein was loaded per sample and 5 ng recombinant E. coli GFP protein (Abcam, ab119740) was loaded as the positive control. Coomassie-stained gel loading control for each time point are shown in lower panels. Imaging of proteins was done on a Bio-Rad Imaging System utilizing Bio-Rad’s Stain Free Technology. Raw gel and blot images included in Additional file 3