The Role of Vascular Actors in Two Dimensional Dialogue of Human Bone Marrow Stromal Cell and Endothelial Cell for Inducing Self-Assembled Network
Figure 2
Expression of VE-cad in HBMSC, HUVEC, co-HBMSC and co-HUVEC after being cultured for different time.
(A) The VE-cad expression was detected by Q-PCR for mRNA level. Data of gene expression was quantified relative to VE-cad gene expressions of HUVEC after 6 h of culture. During 6 hours–18 hours, the gene expression of VE-cad kept constant in HUVEC and co-HUVEC and there is no significant difference of VE-cad gene expression in HUVEC as compared that in co-HUVEC. However, a significant increase of VE-cad gene expression in co-HUVEC was detected at 24 hours while the gene expression of VE-cad in HUVEC still maintained constant. (B) The VE-cad expression was detected by western blot for obtaining protein level. For western blot assay, α-tubulin was analyzed as loading control. The analysis was carried out based on three independent experiments and the gel photos were taken as a representative (not show the other two experiment gel photos). There is no VE-cad detected in co-HBMSC, which indicated the separation of co-cultured cells was completed. a indicated the difference p≤0.05. VE-cad protein expression confirmed the results of gene expression, which showed that there is no significant difference of VE-cad expression between HUVEC and co-HUVEC during 6 hours–18 hours but the co-HUVEC expressed much higher VE-cad than HUVEC at 24 hours. (C) Immunofluorensence staining of VE-cad in co-HUVEC and HUVEC in red. Nuclear was stained in blue with DAPI. Scare bars represent 50 µm. VE-cad started to show at the junction of co-HUVEC from 14 hours and located at the junction of HUVEC all the time.