Neither DUX4 nor DUX4c can induce pitx1 expression. A-B) In situ hybridizations for pitx1 indicate that neither embryos injected with DUX4 nor DUX4c mRNA upregulate pitx1. C) Graphic summary showing none of the embryos tested by in situ hybridization had obvious upregulation of pitx1, as illustrated by percent embryos without increased probe, with numbers of animals assayed. D) Quantification of pitx1 mRNA in DUX4c (1 ng) or DUX4 injected embryos as determined by qRT-PCR. Fold change is normalized to gapdh and compared to EGFP at a relative level of 1; error bars are ±standard error of the mean. Significance (p-value < 0.05) is indicated by (*).