3D structure of MP06 and cell viability. (A) MP06 contains an N-terminal α-helix and unstructured C-terminal region. Basic amino acids contribute to positive net charge of MP06. (B) Electrostatic potential of MP06. Blue and red regions are the positively and negatively charged regions. (C) Hydrophobic and hydrophilic regions of MP06 are visualized as red and white. (D) FITC-MP06 accumulation was observed in the cytoplasm of H1299 cells. Scale bar, 100 μm. (E) Morphological changes from a spindle shape to a cobblestone-like shape in H1299 cells induced by MP06 and FITC-MP06 peptides. Scale bar, 500 μm. (F) Cell viability. ^**P<0.01, ^***P<0.001; vs. control group. MP06, marine-derived peptide 06; con, control.

Regulation of stemness potential and sphere forming ability induced by MP06 in lung cancer cells. (A) Sphere forming assay of H1299 cells treated with MP06. (B) Cell levels of CSC markers in H1299 cells with MP06 using RT PCR and WB. (C) Immunofluorescence staining for expression of SOX2 and FITC-MP06. Scale bar, 100 μm. ^***P<0.001 vs. control. MP06, marine-derived peptide 06; RT, reverse transcription; WB, western blotting; OCT, octamer binding transcription factor; KLF, Kruppel-like factor.

Regulation of EMT and migratory and invasive activity of MP06 in H1299 cells. (A) Wound healing in H1299 cells treated with MP06. (B) Migration and invasion abilities of H1299 cells following treatment with MP06. Scale bar, 500 μm. (C) Cellular levels of EMT markers in H1299 cells treated with MP06. (D) Expression of vimentin and FITC-MP06 through immunofluorescence staining. Scale bar, 100 μm. ^**P<0.01, ^***P<0.001 vs. control. EMT, Epithelial-mesenchymal transition; MP06, marine-derived peptide 06; RT PCR, reverse transcription-PCR; WB, western blotting; ZEB, Zinc-finger E-box-binding homeobox.

Suppression of angiogenesis and expression of VEGF induced by MP06. (A) Cytotoxicity of H1299 cells and HUVECs treated with MP06. (B) Expression of VEGF in H1299 cells treated with MP06. (C) Tube formation ability of HUVECs in conditioned medium (fresh EBM media/H1299 cultured media with MP06 ratios(75:25, 50:50). (D) Formation of tube-like structures in HUVECs reduced by MP06. Scale bar, 500 μm. ^**P<0.01, ^***P<0.001 vs. control. VEGF, vascular endothelial growth factor; HUVEC, Human Umbilical Vein Endothelial Cells; RT, reverse transcription; WB, western blotting; MP06, Marine peptide 06.

MP06-mediated suppression of angiogenesis by MP06 in zebrafish embryos. Lateral view of (A) untreated and (B) MP06-treated embryos exhibit reduced growth of ISVs. (C) Quantitative comparative analysis of the angiogenic effects in the control group and the treated with MP06 group. positive phenotype showing enhanced ISV growth (solid arrows) and the negative phenotype (hollow arrows). (D) Proportion of completed ISV structure in MP06-treated zebrafish at 30 h post-fertilization compared with untreated controls scale bar, 200 μm. ^**P<0.01 vs. control. MP06, Marine peptide 06; ISV, intersegment vessels.

Expression of AQP3 in lung cancer. (A) Levels of AQP3 in lung cancer (H1299) and normal cells (MRC5 and HUVEC). (B) Gene expression analysis of AQP3 in normal and cancer lung tissue using Gene Expression Profiling Interactive Analysis. ^*P<0.05 (C) Kaplan-Meier survival graph of patients with lung cancer according to AQP3 expression. AQP, aquaporin; HUVEC, human umbilical vein endothelial cell.

Regulation of stemness, EMT and angiogenesis by downregulating AQP3. (A) Expression of AQP3 and cancer stem cell markers in AQP3-knockdown H1299 cells. (B) Sphere-forming ability in AQP3-knockdown H1299 cells. Scale bar, 100 μm. (C) Expression of AQP3 and EMT markers in AQP3-knockdown H1299 cells. (D) Migratory and invasive ability of AQP3-knockdown H1299 cells. Scale bar, 200 μm. (E) Angiogenesis activity suppressed by conditioned medium (fresh EBM media/H1299 cultured media with siAQP3 ratios (75:25, 50:50). (F) VEGF expression. ^**P<0.01, ^***P<0.001. EMT, epithelial-mesenchymal transition; AQP, aquaporin; RT, reverse transcription; WB, western blotting; si, small interfering; OCT, octamer binding transcription factor; KLF, Kruppel-like factor; VEGF, vascular endothelial growth factor; RCF, relative colony forming; ZEB, Zinc-finger E-box-binding homeobox.