Fig. 2

Transient expression of G-CSF:eYFP variants in N. benthamiana plants. Expression of Sec-G-CSF:eYFP (a–c), Cyt-G-CSF:eYFP (d–f), CyteYFP (g–i), Zera-G-CSF:eYFP (j–l). White arrows indicate apoplast localization (c) or absence of signal in the apoplast (e, f, h, i). White arrow indicating Golgi localization (b). Inset white boxes (e, f) depict microscopic zoom in shown in f and i, respectively. Samples collected at 4 dpi. Size bar: 5 μm. m Western Blot detection of 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). All samples were treated with PBS + Tween (PBST) extraction buffer and reducing extraction buffer (SDS-DTT). n Band quantification of SDS-DTT treated samples. 20 µg TSP of PBS samples were loaded on the gel. 9 µL (equalized volume to the smallest PBS sample amount) of SDS-DTT samples was loaded on the gel. Black arrows denote monomeric G-CSF:eYFP variants. eGEHK: protein standard. Proteins were detected with GFP antibody. Samples collected at 6 dpi. Columns denoted with a different letter are significantly different (p ≤ 0.05) using one-way ANOVA and followed by Tukey test. Error bars are standard deviation of the means