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Image Search Results
Journal: Molecular Therapy. Nucleic Acids
Article Title: LOXL1 exerts oncogenesis and stimulates angiogenesis through the LOXL1-FBLN5/αvβ3 integrin/FAK-MAPK axis in ICC
doi: 10.1016/j.omtn.2021.01.001
Figure Lengend Snippet: LOXL1 catalytic domain (CD) promotes angiogenesis of VECs by activating the FAK and MAPK signaling pathway (A) Schematic illustration of LOXL1 structure: LOXL1 full-length was divided into five fragments for further purification: F0 (full-length protein), F1 (truncation of signal peptide, spanning residues 26−574 aa), F2 (truncation of both signal and pro-peptides, spanning residues 95−574 aa), F3 (middle fragment, residues 106−574 aa), and F4 (CD, residues 367−574 aa). (B) DNA electrophoresis of five fragments. (C) Purification of GST-LOXL1 CD by passing through a GST column. (D) Identification of GST-LOXL1 CD by HRV 3C protease digestion. (E) Purification of GST-LOXL1 CD by passing through a size-exclusion chromatography column. (F) GST-LOXL1 CD (wild-type [WT]) promoted angiogenesis of VECs compared to GST and GST-LOXL1 CD H449/H451A (mutant). Scale bars, 100 μm (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). (G) pFAK pY861 and pErk1/2 in VECs was upregulated by GST-LOXL1 CD .
Article Snippet: Flag-tag, FBLN5, FAK pY861, FAK pY397, and
Techniques: Purification, Nucleic Acid Electrophoresis, Size-exclusion Chromatography, Mutagenesis
Journal: Molecular Therapy. Nucleic Acids
Article Title: LOXL1 exerts oncogenesis and stimulates angiogenesis through the LOXL1-FBLN5/αvβ3 integrin/FAK-MAPK axis in ICC
doi: 10.1016/j.omtn.2021.01.001
Figure Lengend Snippet: RGD domain in FBLN5 is indispensable to the proangiogenic function of LOXL1 in ICC by binding to the αvβ3 integrin (A) pUC57, pFBLN5 RGD , and pFBLN5 RGE were transfected into RBE cells, and the efficiency of transfection was measured by western blot. A higher level of FBLN5 protein was contained in the supernatant of pFBLN5 transfected groups compared to pUC57 groups. (B and C) The roles of RGD domain and the αvβ3 integrin in the proangiogenic process of GST-LOXL1 CD were detected through tube-formation assays. Cyclo (-RGDfK) (50 ng/μL) was used to block the αvβ3 integrin. Scale bars, 100 μm (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). (D) Cell lysates of VECs were collected for testing phosphorylation levels of FAK and Erk1/2 by western blot. (E) Colocalization of LOXL1, FBLN5, and the αvβ3 integrin along with the vessel-like structure in the extracellular matrix of ICC tumor tissues. Scale bars, 100 μm.
Article Snippet: Flag-tag, FBLN5, FAK pY861, FAK pY397, and
Techniques: Binding Assay, Transfection, Western Blot, Blocking Assay, Phospho-proteomics