GSEA of the top 100 most significantly differentially abundant proteins.

(A) The graph shows enriched terms upon baculovirus infection for the GO category cellular component with a significant depletion of the term ECM. (B) The bar chart presents enriched categories upon baculovirus infection utilizing KEGG pathway as functional database and a significant depletion of the term ECM-receptor interaction (FDR < 0.05).</p

Comparison proteins in EVs.

(XLSX)</p

Protein identification and quantification.

(XLSX)</p

Schematic overview of the employed EV isolation protocol.

Tnms42 cells were grown uninfected and infected with baculovirus at MOI 5 in parallel. EVs were isolated from conditioned media 48 hpi. Several low-speed centrifugation steps eliminated cells, cell debris and large vesicles. A filtration step was applied to additionally clarify the conditioned media. Via ultracentrifugation EVs were first caught in a sucrose cushion, washed, and pelleted via another ultracentrifugation step.</p

Physical characterization of <i>Tnms42</i> cell EVs.

TEM pictures of negative stained EVs from uninfected (A) and infected (B) cells showed cup-shaped vesicles (arrows). Bars 100 nm. Western blot analysis (C) revealed incorporation of EV marker proteins Hsp90 and Hsp70 into Tnms42 cell EVs and an enrichment of Hsp90 in EVs upon baculovirus infection.</p

KEGG pathway analysis.

(DOCX)</p

Proteomic analysis of EVs from uninfected and baculovirus infected <i>Tnms42</i> cells.

(A) The volcano plot shows differentially abundant proteins in EVs from infected compared to uninfected cells. The horizontal dashed grey line represents the set significance threshold (> 15) and significantly differentially abundant proteins are highlighted in black. (B) The bar chart shows the KEGG pathway analysis and the number of matched objects for the top 15 categories with the most matched objects, including metabolic pathways, endocytosis and signalling pathways.</p

S1 Raw images -

(PDF)</p

Glycosylation profile of recombinant HAs from leaves and seeds.

<p>Man_{6, 7, 8}: Mannose _{6, 7, 8}; Gn: <i>N</i>-acetylglucosamine; X: Xylose; F: Fucose.</p

Purification of ELPylated hemagglutinin (H5-ELP) from transgenic leaves and seeds by membrane-based ITC.

<p>ELPylated hemagglutinins from leaves (A) and seeds (B) were purified by the standard or improved mITC methods (C) described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099347#s2" target="_blank">Materials and Methods</a> section. Proteins in the raw plant extract (RE), in the supernatant after passage through a 0.2 µm cellulose acetate membrane (Sm) and in the eluent (Pm) were collected during the mITC purification process and separated by 10% SDS-PAGE. Recombinant proteins were then detected using Coomassie Brilliant Blue staining (left) or an anti-c-myc monoclonal antibody (right).</p