Fig. 6

Effect of pharmacological Notch signaling inhibition and activation on gene transcription in 4.0 dpc mouse expanded blastocysts. Mouse 2.5 dpc embryos were in vitro cultured in the presence of a Notch inhibitor (DAPT) or of Notch ligands Jagged1 and Jagged2, for 36 h, until 4.0 dpc. Expanded blastocysts from groups Control (n = 5), Jagged1-treated (n = 5), Jagged2-treated (n = 5) and DAPT-treated (n = 6) were processed for qRT-PCR analysis. a-j: Transcription of Notch1 (a), Notch2 (b), Jagged1 (c), Jagged2 (d) and Hes1 (e), and of pluripotency and differentiation genes Sox2 (f), Oct4 (g), Klf4 (h), Cdx2 (i) and Cdca7 (j) were analyzed. Bars represent Log₂ of power of ∆∆Ct values. These values were generated by first normalizing the Ct values of each target gene with the mean Ct values of the endogenous control genes Rps29 and Hprt1. The obtained ∆Ct values were then calibrated to ∆Ct values of Control embryos, which were used as calibrators (shown as 0.0), originating the ∆∆Ct values for log transformation; error bars show the standard error of the mean (s.e.m). Exact ANOVA results (p) are shown for each gene. Different letters within the same gene represent significantly different mean values (p < 0.05; LSD post-hoc). *Transcription of Jagged1 differs significantly (p = 0.038) between groups Control and Jagged2-treated (T-test). k: Agarose gels of qRT-PCR products of genes Jagged1, Jagged2 and Cdx2. Images illustrate results from representative 4.0 dpc expanded blastocysts from groups Control (n = 4), Jagged1-treated (n = 5), Jagged2-treated (n = 5) and DAPT-treated (n = 5). The DNA ladder has 50 bp increments, and the arrow (→) signals the 50 bp mark