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. 2013 Apr 17;8(4):e61506. doi: 10.1371/journal.pone.0061506

Figure 1. Effects of Thy-1 over-expression on proliferation, migration and capillary-like tube formation in vascular endothelial cells.

Figure 1

(a) Representative micrographs of EGFP-positive cells in bright-field image (right panel) and immunofluorescence (left panel) are indicated by the arrows. (b) Quantitative results of the HUVEC expressing Thy-1-pCMS-EGFP-C1 (n = 3). The transfection efficiency of Thy-1-positive cells is around 40%. Transfection of the HUVEC with Thy-1-pcDNA3.1 caused an increase of the levels of Thy-1 mRNA (c) and protein (d) as detected by RT-PCR and Western blot analyses, respectively. For Western blot analysis, membrane was probed with anti-G3PDH antibody to verify equivalent sample loading. RT-PCR products of G3PDH were used as an internal control for RT-PCR analyses. (e) Over-expression of Thy-1 did not affect the proliferation of HUVEC. The cells were transfected with pcDNA301 (control) or Thy-1 pcDNA3.1 for 96 h and then harvested for MTT assay. Values represent the means±s.e.mean. (n = 5). Over-expression of Thy-1 inhibited the migration of HUVEC (f) and HMEC-1 (g). Top panel: a representative result of the migrated HUVEC transfected with pcDNA301 or Thy-1 pcDNA3.1. Bottom panel: quantification of migration inhibition. The quantified results were expressed by percentage of control. Values represent the means±s.e.mean. (n = 3). (h) Over-expression of Thy-1 inhibited the capillary-like tube formation.