MOESM2 of Transient co-expression with three O-glycosylation enzymes allows production of GalNAc-O-glycosylated Granulocyte-Colony Stimulating Factor in N. benthamiana (original) (raw)

Additional file 2: Figure S2. Secretory and Cytoplasmic G-CSF can be extracted with a PBS-based extraction buffer, but Zera-G-CSF can only be extracted under denaturing and reducing conditions. Western Blot detection of G-CSF:eYFP variants transiently expressed in N. benthamiana plants. Samples collected at 4 dpi. Upper panel: Plant 1; Lower panel: Plant 2. Sec-G-CSF:eYFP (Secretory variant, 49 kDa), Cyt-G-CSF:eYFP (Cytoplasmic and mature protein variant, 49 kDa), Zera-G-CSF:eYFP (Zera fused variant, 61 kDa), eYFP (Cytoplasmic eYFP, 29 kDa), p19 negative control. Four leaf discs from two different plants were collected for each sample. 50 µg TSP of PBST samples or equivalent volume of SDS-DTT samples were loaded on the gel. Black arrows denote Zera-G-CSF:eYFP. Gray arrows denote Sec-GCSF-:eYFP and Cyt-G-CSF:eYFP. Red asterisks denotes a faint band corresponding to Zera-G-CSF:eYFP extracted with PBST extraction buffer and its corresponding proper extraction with SDS-DTT extraction bu...

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