Fig. 3

Overexpression of GLI2 does not significantly upregulate the number of MyHC^+ve cells. a MyHC^+ve cells (red) were visualized and (b) counted in corresponding day 7 differentiating cells by indirect immunofluorescence. Hoechst (blue) was used to visualize nuclei. Representative images of the cardiomyocyte-enriched areas on the periphery of an EB are shown. Scale bar represents 100 μm. At least 2500 nuclei were counted across 20 random fields of view, per biological replicate; n = 3. c Myhc6/7 expression levels on the days indicated in mES[Ctrl] (white bars) and mES[Gli2] (grey bars) cells were normalized to β-actin, calibrated to day 0 mES[Ctrl] culture expression levels, and presented as a percentage of the highest expression level recorded, per gene. Error bars represent +/- SEM; n = 3. One-tailed Student’s T-tests were used for statistical analyses. Grey lines represent paired T-tests; black lines represent unpaired T-tests; ( ) p < 0.05. d MyHC^+ve cells from (a) were analyzed for MyHC signal intensity using the ImageJ program. At least 100 cells were analyzed across 10 random fields of view per biological replicate; n = 2