Figure 5

Ectopic expression of Syndecan-2 and -4 through differentiation of CCE cells to mesodermal lineages. Undifferentiated CCE cells (A) as well as Flk1⁺ (B) and Flk1^- (C) cells FACS sorted on day 4 of differentiation were analysed by RT-PCR for syndecan expression. D. CCE cells were transfected with either pCAGIPuro-sdc2 or pCAGIPuro-sdc4 and were selected for puromycin resistance. FACS analysis was performed on undifferentiated and cells differentiated to the Flk1 stage as indicated using antibodies against syndecan-2 and -4. In each analysis, the dotted black line denotes the relevant secondary antibody control, the blue line corresponds to cells transfected with pCAGIPuro empty vector and the red line are cells transfected with constructs containing syndecan-2 or-4 cDNA. E. Levels of Flk1 were measured on day 5 of differentiation in cells transfected with pCAGIPuro-sdc2, pCAGIPuro-sdc4 or empty vector control. Data from 5 independent experiments are shown, where a total of 9 flasks were analysed for each construct. The bars represent the standard deviation, and the results were analysed with repeated-measures ANNOVA, using Prism Graphpad 4.0, assuming Gaussian distribution. No statistical significance was observed for the difference in FLK1 expression in cells transfected with the different constructs.