Additional file 1 of C-terminal mini-PEGylation of a marine peptide N6 had potent antibacterial and anti-inflammatory properties against Escherichia coli and Salmonella strains in vitro and in vivo
Creators: name: Li, Ting
name: Yang, Na
name: Teng, Da
name: Mao, Ruoyu
name: Hao, Ya
name: Wang, Xiumin
name: Wang, Jianhua
Publisher: name: figshare
Year: 2022
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Figure S1. Chemicalstructure of N6. Figure S2. Chemicalstructure of N6-COOH-miniPEG. Figure S3. Chemical structure of N6-NH2-miniPEG.Figure S4. Chemical structure of N6-NH2-PEG6. Figure S5. Chemicalstructure of N6-NH2-PEG12. Figure S6. Chemical structure of N6-NH2-PEG24. Figure S7. Chemicalstructure of N6-Cys7-miniPEG. Figure S8. Chemicalstructure of N6-Cys16-miniPEG. Figure S9. CD of N6 and N6-COOH-miniPEG indifferent solutions. Figure S10. Interaction of N6 and N6-COOH-miniPEG with cell membrane. Outer membrane permeabilization of E. coli CVCC195(A) and S. pullorum CVCC533 (B) cells after treated with N6and N6-COOH-miniPEG. Figure S11. Interactionof N6 and N6-COOH-miniPEGwith cell membrane. Inner membranepermeabilization of E. coli CVCC195 (A) and S. pullorum CVCC533 (B) cells. Bacterial cells were treated with 1×, 2× or 4× MIC for 5, 30 or 120 minand analyzed by flow cytometry. Figure S12. Interactionof N6 and N6-COOH-miniPEGwith cell membrane. (A-D) Effects of N6 and N6-COOH-miniPEG on E. coli CVCC195 (A,B) and S. pullorum CVCC533 (C, D) cytoplasmicmembrane potential. Figure S13. Binding affinity of N6 andN6-COOH-miniPEG to LPS. Figure S14. TEM images of E. coli CVCC195 and S. pullorumCVCC533 cells treated with N6 and N6-COOH-miniPEG. After treatment with 4 × MIC N6 or N6-COOH-miniPEGfor 2 h, E. coli CVCC195 and S. pullorum CVCC533 cells weredehydrated, sputtered, and observed on JEM1400 (JEDL, Tokyo, Japan). Figure S15. Effects of N6 and its N6-COOH-miniPEG on organ injury in mice.The mice were infectedintraperitoneally with E. coli CVCC195 (1×109 CFU/mL, 200 μL)and treated with N6 (4 μmol/kg) or N6-COOH-PEG (4 μmol/kg). The livers,spleens, kidneys and lungs were harvested from the mice sacrificed at 5 d afterinfection. Figure S16. Effectsof N6 and its N6-COOH-miniPEG on organ injury in mice. The mice wereinfected intraperitoneally with S. pullorum CVCC533 (5 × 107CFU/mL, 200 μL) and treated with N6 (10 μmol/kg) or N6-COOH-PEG (10 μmol/kg).The livers, spleens, kidneys and lungs were harvested from the mice sacrificedat 48 h after infection. Figure S17. Effects ofN6 and N6-COOH-miniPEGon organ injury in mice.Mice were infectedintraperitoneally with S. pullorum CVCC533 (5 × 107 CFU/mL,200 μL) and treated with N6 (10 μmol/kg) or N6-COOH-miniPEG (10 μmol/kg). Livers, spleens, kidneys and lungs were harvested from mice sacrificed at 4 dafter infection. CK group: The livers, spleens, kidneys, and lungs were normal; Infected S. pullorum CVCC533 group: Thereis “bridgingnecrosis” around the confluent area of the liver, a marked decrease in the density oflymphocytes in the splenic nodes, diffuse inflammatory cell infiltration in therenal interstitium, “honeycomb” changes throughout the lung tissue, and a large amount of bloody exudate in thealveolar cavity (arrow) (× 200,scale bar = 100 μm); N6 treatment group: Foci of hepatocyte necrosis are seenin the lobules of the liver, localised reduced lymphocyte density in thesplenic nodules, localised renal tubular degeneration and atrophy, and a smallamount of inflammatory cell infiltration in the interstitial lung (arrow) (× 200, scale bar = 100 μm); N6-COOH-miniPEG treatment group: Focal foci of hepatocyte necrosis are seen inthe lobules of the liver, localised reduced lymphocyte density in the splenicnodules, localised renal tubular degeneration and atrophy, and a small amountof scattered inflammatory cell infiltration in the interstitial lung (× 200, scale bar = 100 μm); CIP treatment group: Localized inflammatory cell infiltration in the hepaticsinusoids, reduced lymphocyte density in the splenic nodules, localizedscattered inflammatory cell infiltration in the renal interstitium, markedwidening of the alveolar septa and narrowing of the alveolar cavity (× 200, scale bar = 100 μm). Table S1. Proportion of secondary structure of N6 and N6-COOH-miniPEG in different solutions. Table S2. MIC values (μg/mL) of N6 and N6-COOH-miniPEG against E. coli CVCC195 in differentconditions.