The changes in mRNA levels were analyzed using the Bio-Rad iQ5 2.1 Standard Edition Optical System Software. Undifferentiated cells (day 0) were used as a calibrator, and mean Ct values for the target genes were normalized against the PPIA housekeeping gene. The results obtained from the quantitative real-time PCR experiments were calculated using the 2-ΔΔCt method. The intensity of the protein bands obtained from the western blot experiments was quantitated using ImageJ (1.48 version). The results were compared to undifferentiated cells and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control.
The changes in gene expression and protein amounts were statistically analyzed with the non-parametric Kruskal-Wallis test, using the GraphPad Prism (6.01) software. Values p < 0.05 were considered significant. The time points of differentiation (days 3, 5, 7) were compared to the undifferentiated state (day 0) with Dunn’s multiple comparison test. Graphics were drawn with mean values using the GraphPad Prism (6.01) software with error bars equivalent to the standard error of the mean (SEM).
The changes in gene expression and protein amounts were statistically analyzed with the non-parametric Kruskal-Wallis test, using the GraphPad Prism (6.01) software. Values p < 0.05 were considered significant. The time points of differentiation (days 3, 5, 7) were compared to the undifferentiated state (day 0) with Dunn’s multiple comparison test. Graphics were drawn with mean values using the GraphPad Prism (6.01) software with error bars equivalent to the standard error of the mean (SEM).