Figure 4.
Knock-down MBF-1 function by microinjection of a synthetic mRNA encoding for as dominant repressor (dnMBF-1). Increasing amounts (0.1–1 pg) of the chimeric RNA were injected in P. lividus zygotes and RNA extracted from embryos at morula stage. Graphs show n-fold changes in mRNA expression level of histone genes based on the threshold cycle number (Ct) of dnMBF-1 injected embryos compared to that of the uninjected control embryos. Ct numbers were normalized for the endogenous MBF-1 in the same sample, by amplifying a fragment of the coding region external to the DNA binding domain. Data were derived from two independent microinjection experiments and each bar represents the average of triplicate samples from the two batches of embryos.