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Supplementary File 1 Supplemental Materials GFP DNA GFP DNA Figure S1. Fluorescence microscopy images of GFP-labelled hCECs cultured in 2D monocultures shows high GFP expression. F-actin F-actin PKH26 PKH26 DNA DNA F-actin PKH26 DNA Figure S2. PKH26-stained hCFs show broad cytosolic fluorescent staining. Relatively equal distribution of PKH26 throughout the culture shown in low (left side) and high magnification images (right side). Cells 2019, 8, x; doi: FOR PEER REVIEW www.mdpi.com/journal/cells Cells 2019, 8, x FOR PEER REVIEW 2 of 21 F-actin F-actin CD63 CD9 DNA DNA Figure S3. Expression of exosomal markers, CD63 and CD9, in hCECs cultured in 2D monocultures. Figure S4. Representative brightfield images of the collagen contraction assay. Cells 2019, 8, x FOR PEER REVIEW 3 of 21 Table S1. Proteins identified in isolated epithelial-derived (hCE-TJ) EVs categorized by dominant subcellular origin. Proteins are classified based on nine major subcellular locations: 1) Cytoplasm (Soluble), 2) Cytoplasm (Cytoskeleton/Motor), 3) Cytoplasm (Endosomal Trafficking), 4) Extracellular (Secreted), 5) Plasma Membrane, 6) Mitochondria, 7) Nucleus, 8) ER/Golgi, and 9) Other (Lysosome, Unknown). Classification is based on subcellular localization information described in the the UniProtKB database according to the dominant location based on the epithelial cell type (Ref. Breuza L., et al. Database (Oxford). 2016 Feb 20;2016. pii: bav120). MW Protein Name Accession Localization # Peptides [kDa] 1-phosphatidylinositol 4,5-bisphosphate Q4KWH8 Cytoplasm (Soluble) 1 189.1 phosphodiesterase 1,4-alpha-glucan-branching enzyme Q04446 Cytoplasm (Soluble) 6 80.4 3-hydroxybutyrate dehydrogenase type 2 Q9BUT1 Cytoplasm (Soluble) 1 26.7 3-phosphoinositide-dependent protein kinase 1 O15530 Cytoplasm (Soluble) 1 63.1 3'(2'),5'-bisphosphate nucleotidase 1 O95861 Cytoplasm (Soluble) 1 33.4 4-trimethylaminobutyraldehyde dehydrogenase P49189 Cytoplasm (Soluble) 5 53.8 6-phosphogluconolactonase O95336 Cytoplasm (Soluble) 2 27.5 6-phosphogluconate dehydrogenase P52209 Cytoplasm (Soluble) 6 53.1 14-3-3 protein gamma P61981 Cytoplasm (Soluble) 7 28.3 14-3-3 protein epsilon P62258 Cytoplasm (Soluble) 3 29.2 14-3-3 protein eta Q04917 Cytoplasm (Soluble) 5 28.2 14-3-3 protein theta P27348 Cytoplasm (Soluble) 12 27.7 14-3-3 protein zeta/delta P63104 Cytoplasm (Soluble) 6 27.7 26S proteasome non-ATPase regulatory subunit 2 Q13200 Cytoplasm (Soluble) 3 100.1 26S proteasome non-ATPase regulatory subunit 3 O43242 Cytoplasm (Soluble) 4 60.9 26S protease regulatory subunit 4 P62191 Cytoplasm (Soluble) 5 49.2 26S protease regulatory subunit 6A P17980 Cytoplasm (Soluble) 2 49.2 26S protease regulatory subunit 6B P43686 Cytoplasm (Soluble) 3 47.3 26S protease regulatory subunit 7 P35998 Cytoplasm (Soluble) 5 48.6 26S proteasome non-ATPase regulatory subunit 8 P48556 Cytoplasm (Soluble) 2 39.6 26S protease regulatory subunit 10B P62333 Cytoplasm (Soluble) 2 44.1 26S proteasome non-ATPase regulatory subunit 11 O00231 Cytoplasm (Soluble) 2 47.4 26S proteasome non-ATPase regulatory subunit 12 O00232 Cytoplasm (Soluble) 4 52.9 26S proteasome non-ATPase regulatory subunit 13 Q9UNM6 Cytoplasm (Soluble) 4 42.9 26S proteasome non-ATPase regulatory subunit 14 O00487 Cytoplasm (Soluble) 1 34.6 40S ribosomal protein S2 P15880 Cytoplasm (Soluble) 8 31.3 40S ribosomal protein SA P08865 Cytoplasm (Soluble) 3 32.8 40S ribosomal protein S3 P23396 Cytoplasm (Soluble) 12 26.7 40S ribosomal protein S3a P61247 Cytoplasm (Soluble) 5 29.9 40S ribosomal protein S4, X isoform P62701 Cytoplasm (Soluble) 9 29.6 40S ribosomal protein S5 P46782 Cytoplasm (Soluble) 2 22.9 40S ribosomal protein S7 P62081 Cytoplasm (Soluble) 1 22.1 Cells 2019, 8, x FOR PEER REVIEW 4 of 21 40S ribosomal protein S8 P62241 Cytoplasm (Soluble) 1 24.2 40S ribosomal protein S9 P46781 Cytoplasm (Soluble) 6 22.6 40S ribosomal protein S11 P62280 Cytoplasm (Soluble) 3 18.4 40S ribosomal protein S13 P62277 Cytoplasm (Soluble) 5 17.2 40S ribosomal protein S14 P62263 Cytoplasm (Soluble) 1 16.3 40S ribosomal protein S15a P62244 Cytoplasm (Soluble) 2 14.8 40S ribosomal protein S16 P62249 Cytoplasm (Soluble) 5 16.4 40S ribosomal protein S18 P62269 Cytoplasm (Soluble) 1 17.7 40S ribosomal protein S19 P39019 Cytoplasm (Soluble) 1 16.1 40S ribosomal protein S25 P62851 Cytoplasm (Soluble) 3 13.7 40S ribosomal protein S26 P62854 Cytoplasm (Soluble) 1 13 40S ribosomal protein S27-like Q71UM5 Cytoplasm (Soluble) 1 9.5 60S ribosomal protein L4 P36578 Cytoplasm (Soluble) 2 47.7 60S ribosomal protein L5 P46777 Cytoplasm (Soluble) 3 34.3 60S ribosomal protein L7 P18124 Cytoplasm (Soluble) 2 29.2 60S ribosomal protein L9 P32969 Cytoplasm (Soluble) 1 21.9 60S ribosomal protein L10a P62906 Cytoplasm (Soluble) 1 24.8 60S ribosomal protein L11 P62913 Cytoplasm (Soluble) 3 20.2 60S ribosomal protein L12 P30050 Cytoplasm (Soluble) 2 17.8 60S ribosomal protein L13 P26373 Cytoplasm (Soluble) 1 24.2 60S ribosomal protein L22 P35268 Cytoplasm (Soluble) 1 14.8 60S ribosomal protein L23 P62829 Cytoplasm (Soluble) 1 14.9 60S ribosomal protein L30 P62888 Cytoplasm (Soluble) 1 12.8 60S ribosomal protein L34 P49207 Cytoplasm (Soluble) 2 13.3 60S ribosomal protein L35 P42766 Cytoplasm (Soluble) 1 14.5 60S ribosomal protein L36 Q9Y3U8 Cytoplasm (Soluble) 1 12.2 60S acidic ribosomal protein P0 P05388 Cytoplasm (Soluble) 4 34.3 60S acidic ribosomal protein P2 P05387 Cytoplasm (Soluble) 1 11.7 Acyl-protein thioesterase 2 O95372 Cytoplasm (Soluble) 1 24.7 Adenylate kinase isoenzyme 1 P00568 Cytoplasm (Soluble) 2 21.6 Adenosylhomocysteinase P23526 Cytoplasm (Soluble) 9 47.7 Adenylyl cyclase-associated protein 1 Q01518 Cytoplasm (Soluble) 1 51.9 ADP-ribosylation factor 3 P61204 Cytoplasm (Soluble) 4 20.6 Adenosine kinase P55263 Cytoplasm (Soluble) 2 40.5 Adenosylhomocysteinase 2 O43865 Cytoplasm (Soluble) 6 58.9 Alanine-tRNA ligase P49588 Cytoplasm (Soluble) 3 106.7 Alcohol dehydrogenase class-3 P11766 Cytoplasm (Soluble) 5 39.7 Alcohol dehydrogenase P14550 Cytoplasm (Soluble) 2 36.6 Aldehyde dehydrogenase family 1 member A3 P47895 Cytoplasm (Soluble) 4 56.1 Aldose 1-epimerase Q96C23 Cytoplasm (Soluble) 1 37.7 Aldose reductase P15121 Cytoplasm (Soluble) 5 35.8 Alpha-aminoadipic semialdehyde dehydrogenase P49419 Cytoplasm (Soluble) 5 58.5 Cells 2019, 8, x FOR PEER REVIEW 5 of 21 Alpha-enolase P06733 Cytoplasm (Soluble) 8 47.1 Aminoacyl tRNA synthase complex Q12904 Cytoplasm (Soluble) 1 34.3 AMP deaminase 2 Q01433 Cytoplasm (Soluble) 2 100.6 Asparagine-tRNA ligase O43776 Cytoplasm (Soluble) 1 62.9 Aspartyl aminopeptidase Q9ULA0 Cytoplasm (Soluble) 1 52.4 Aspartate aminotransferase P17174 Cytoplasm (Soluble) 4 46.2 Aspartate-tRNA ligase P14868 Cytoplasm (Soluble) 2 57.1 ATP-citrate synthase P53396 Cytoplasm (Soluble) 8 120.8 ATP-dependent RNA helicase DDX1 Q92499 Cytoplasm (Soluble) 3 82.4 Basic leucine zipper and W2 domain-containing Q7L1Q6 Cytoplasm (Soluble) 1 48 protein 1 Bifunctional glutamate/proline--tRNA ligase P07814 Cytoplasm (Soluble) 3 170.5 Bifunctional purine biosynthesis protein PURH P31939 Cytoplasm (Soluble) 6 64.6 Bleomycin hydrolase Q13867 Cytoplasm (Soluble) 1 52.5 C-1-tetrahydrofolate synthase, cytoplasmic P11586 Cytoplasm (Soluble) 5 101.5 C-terminal-binding protein 1 Q13363 Cytoplasm (Soluble) 2 47.5 Calcyclin-binding protein Q9HB71 Cytoplasm (Soluble) 1 26.2 Calreticulin P27797 Cytoplasm (Soluble) 1 48.1 cAMP-dependent protein kinase catalytic subunit P22694 Cytoplasm (Soluble) 1 40.6 beta cGMP-dependent protein kinase 1 Q13976 Cytoplasm (Soluble) 1 76.3 Coatomer subunit beta' P35606 Cytoplasm (Soluble) 2 102.4 Complement C3 P01024 Cytoplasm (Soluble) 5 187 Complement factor I P05156 Cytoplasm (Soluble) 3 65.7 COP9 signalosome complex subunit 1 Q13098 Cytoplasm (Soluble) 3 55.5 COP9 signalosome complex subunit 2 P61201 Cytoplasm (Soluble) 2 51.6 COP9 signalosome complex subunit 4 Q9BT78 Cytoplasm (Soluble) 3 46.2 COP9 signalosome complex subunit 5 Q92905 Cytoplasm (Soluble) 2 37.6 CTP synthase 2 Q9NRF8 Cytoplasm (Soluble) 1 65.6 Cullin-associated NEDD8-dissociated protein 1 Q86VP6 Cytoplasm (Soluble) 11 136.3 Cytoplasm aminopeptidase P28838 Cytoplasm (Soluble) 2 56.1 D-3-phosphoglycerate dehydrogenase O43175 Cytoplasm (Soluble) 3 56.6 Dihydropteridine reductase P09417 Cytoplasm (Soluble) 1 25.8 Dihydropyrimidinase-related protein 2 Q16555 Cytoplasm (Soluble) 11 62.3 Dihydropyrimidinase-related protein 3 Q14195 Cytoplasm (Soluble) 7 61.9 Dihydropyrimidinase-related protein 5 Q9BPU6 Cytoplasm (Soluble) 3 61.4 DNA damage-binding protein 1 Q16531 Cytoplasm (Soluble) 5 126.9 DnaJ homolog subfamily B member 1 P25685 Cytoplasm (Soluble) 1 38 E3 ubiquitin-protein ligase HUWE1 Q7Z6Z7 Cytoplasm (Soluble) 2 481.6 E3 ubiquitin-protein ligase RNF213 Q63HN8 Cytoplasm (Soluble) 1 591 Elongation factor 1-alpha 1 P68104 Cytoplasm (Soluble) 12 50.1 Elongation factor 1-gamma P26641 Cytoplasm (Soluble) 9 50.1 Cells 2019, 8, x FOR PEER REVIEW 6 of 21 Elongation factor 2 P13639 Cytoplasm (Soluble) 19 95.3 Eukaryotic initiation factor 4A-II Q14240 Cytoplasm (Soluble) 5 46.4 Eukaryotic peptide chain release factor P15170 Cytoplasm (Soluble) 2 55.7 Eukaryotic translation initiation factor 2 subunit 1 P05198 Cytoplasm (Soluble) 4 36.1 Eukaryotic translation initiation factor 2 subunit 3 P41091 Cytoplasm (Soluble) 2 51.1 Eukaryotic translation initiation factor 3 B5ME19 Cytoplasm (Soluble) 3 105.4 Eukaryotic translation initiation factor 3 subunit A Q14152 Cytoplasm (Soluble) 1 166.5 Eukaryotic translation initiation factor 3 subunit D O15371 Cytoplasm (Soluble) 1 63.9 Eukaryotic translation initiation
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