Figure 2. Helja secretion is not affected by brefeldin A treatment. Sunflower seeds were imbibed in water (Control) or 50 μg ml⁻¹ brefeldin A (BFA) for 2 h and subjected to extracellular fluid (EF) extraction or extracellular vesicles preparation as previously described.¹² The EF (5 g of seeds, A) or the vesicles (15 g of seeds, B) were fractionated by SDS-PAGE and the proteins were stained with Coomassie blue (upper panel) or transferred to nitrocellulose for immunodetection of Helja (lower panel). The molecular weight markers are indicated on the left.