Figure 3
EpCAM expression in differentiating ESC. (a) Schematic depiction of the timeline of EB formation. (b) Representative pictures of E14TG2α ESC in 2D culture (ES cells) and embryoid bodies (EB) at the indicated time points of spontaneous 3D-differentiation. (c) Representative FACS histogram of EpCAM expression in pluripotent E14TG2α ESC and EB at differentiation day 21. (d) Mean EpCAM and SSEA1 cell surface expression measured by FACS analysis in pluripotent E14TG2α ESC and EB (d21) (n = 3 independent experiments). (e) Mean EpCAM mRNA expression measured by quantitative PCR in pluripotent E14TG2α ESC and differentiated EB (day 21) (n = 3 independent experiments). (f) Kinetic of EpCAM and Oct3/4 mean mRNA expression measured by quantitative PCR in pluripotent and differentiating ESC (n = 3 independent experiments). (g) Schematic depiction of primer pairs relative to transcription start site (ATG) of EPCAM. (h) Time and concentration kinetics of input control at EPCAM promoter and CENPI locus from chromatin-IP samples. (n = 3 independent experiments). (i) Chromatin-IP (ChIP) of polymerase II (Pol II), H3K4 and H3K27 at EPCAM promoter and CENPI locus (n = 3 independent experiments). Shown are mean values of quantitative PCR amplification of the region of the EPCAM promoter after ChIP with the indicated specific antibodies. (n = 3 independent experiments). Mean ± SEM; Student’s T-test (n = 2 groups) or One-Way ANOVA (n ≥ 3 groups); p < 0.05, **p < 0.01, ***p < 0.001.
