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. 2014 Apr 21;9(4):e95434. doi: 10.1371/journal.pone.0095434

Figure 3. Addition of PD-0325901 inhibited Erk phosphorylation and resulted in more circular embryoid bodies.

Figure 3

Embryoid bodies were grown in different concentrations of PD-0325901 or 0.04% DMSO for 7 days. (A) Light microscopy images show morphology of the embryoid bodies following inhibitor treatment. The embryoid bodies appeared slightly smaller and rounder. Scale bars 200 µm. (B) Western blotting demonstrates that PD-0325901 reduced levels of diphosphorylated Erk1/2. A representative blot and quantification from 3 independent experiments is shown. (C) Quantification of the size of the embryoid bodies suggests that there was a slight reduction in size, but this is not statistically significant. (D) Measurement of the circularity of the embryoid bodies shows that inhibition of Mek caused a statistically significant increase in circularity. (E) Whole-mount immunostaining of cleaved Caspase-3. A slight reduction in the number of cleaved Caspase-3 nuclei is observed upon treatment with 4 µM PD-0325901 in comparison to 0.04% DMSO. A representative image from 3 independent experiments is shown. Data is from 3 independent experiments, error bars are SEM. Statistical analysis is (B–D) a one-way Anova with a Dunnett’s post-hoc test, (E) a paired t-test. (*P = 0.1–0.5, **p = 0.001–0.01, ***p<0.001).