Fig. 3

Control of ZEB1 and Annexin A5b Expression by miRNAs and Dnd(A) Lack of miRNAs in MZdicer embryos results in higher GFP signal from a reporter containing the 3′ UTR of zeb1, measured at 16 hpf from the line marked areas (Figure S2A).(B) qPCR analysis comparing the endogenous levels of zeb1 mRNA in MZdicer and wild-type embryos.(C) The 3′ UTR of zeb1 contains a seed sequence of miR-430 (blue). Binding site for the target protector (TP) morpholino masking the miRNA seed (TP) is indicated and the point mutation disrupting the seed is labeled in red.(D) Application of TPs increases GFP expression from the reporter RNA as compared to embryos treated with control morpholinos.(E) Quantitative representation of the results of the experiment in (D) measured within the area marked by the dotted lines in (D), depicting the GFP signal level normalized to that of a coinjected mCherry control.(F) Expression of the Venus protein from RNA containing the wild-type zeb1 3′ UTR is reduced in dnd MO treated PGCs as compared to that from an RNA reporter mutated for miRNA binding (cfp-3′zeb1 miRmut).(G) Quantitative representation of the results in (F).(H) qPCR analysis of endogenous annexin A5b, nanos, mlck, and zeb1 mRNA in PGCs isolated from Dnd knocked-down embryos compared to control morpholino-injected animals.n is the number of embryos analyzed in (E) or the number of PGCs examined in (G). Error bars depict the SEM and p < 0.001, calculated using two-tailed, unpaired t test.See also Figure S2.