Research Paper Volume 12, Issue 5 pp 4506—4526

LncRNA GUSBP5-AS promotes EPC migration and angiogenesis and deep vein thrombosis resolution by regulating FGF2 and MMP2/9 through the miR-223-3p/FOXO1/Akt pathway

Figure 2. Effects of lncRNA GUSBP5-AS on angiogenesis of EPCs in vivo and in vitro, EPC migration and invasion. (A) In vivo angiogenesis was evaluated at Day 7 after subcutaneous injection of Matrigel-mixed EPCs into nude mice. (B) HE staining: lncRNA GUSBP5-AS knockdown and overexpression, respectively, decreased and increased tube formation by EPCs in vivo (original magnification, ×200). (C) In vitro tube formation assay. Scale bar=100μm. (original magnification, ×100). LncRNA GUSBP5-AS knockdown and overexpression, respectively, decreased and increased tube number. ***P< .001. (D) Wound healing assay showing the effects of lncRNA GUSBP5-AS on EPC migration. Scale bar=100μm. (original magnification, ×100). LncRNA GUSBP5-AS knockdown and overexpression, respectively, decreased and increased cell migration in EPCs. **P< .01. (E) Effects of lncRNA GUSBP5-AS on cell invasion ability were analysed. Scale bar=100μm. (original magnification, ×200). Data are represented as mean ± SEM. ***P< .001. (F) Effect of lncRNA GUSBP5-AS on actin cytoskeleton structure in cultured EPCs. Cells were fixed, permeabilized, stained with rhodamine–phalloidin and DAPI, and visualized by confocal microscopy; down-regulation of GUSBP5-AS impaired F-actin while GUSBP5-AS overexpression prevented disruption of actin cytoskeletal structure.